Journal of Environmental Management 248 (2019) 109312

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Journal of Environmental Management

journal homepage: www.elsevier.com/locate/jenvman

Research article

Enzymatic profiles associated with the evolution of the lignocellulosic T

fraction during industrial-scale composting of anthropogenic waste:
Comparative analysis
M.J. Estrella-González, M.M. Jurado, F. Suárez-Estrella*, M.J. López, J.A. López-González,
A. Siles-Castellano, J. Moreno
Department of Biology and Geology, CITE II-B, University of Almería, Agrifood Campus of International Excellence, CeiA3, 04120, Almería, Spain

ARTICLE INFO ABSTRACT

Keywords: In the new European Waste Law, composting is proposed as one of the best options to properly manage organic
Industrial-scale composting waste of anthropogenic origin. Currently, the massive generation of this type of waste, as well as its hetero-
Lignocellulose geneity, makes difficult in many cases control this process of degradation on an industrial scale.
Humification In this work, 15 facilities were selected based on 5 types of organic waste: Urban Solid Waste, Vegetable
Carbon cycle-related enzymes
Waste, Sewage Sludges, Agrifood Waste and “Alpeorujo”. The samples were collected in different thermal
Compost quality
phases. The results revealed very different physicochemical and enzymatic profiles, as well as different degrees
of humification depending on the process and the raw materials. However, parameters such as β-glucosidase,
amylase, lignin/holocellulose ratio and humification rate showed similar trends in all cases. All of them could act
as important indicators to evaluate the quality of a composting process, despite the heterogeneity of the starting
materials.

1. Introduction properties: a minimum of 35–40% organic matter, no more than 40%

humidity and C/N ratio values below 15–20. Additionally, several
In recent decades, the amount of municipal waste and sludge from parameters have been established to determine the state of maturity of
wastewater treatment facilities has increased significantly due to the a compost, among which are temperature, odor, color, cation exchange
exponential growth of the world population and the consumer habits of capacity, humified fractions and other respirometric variables (Iglesias-
the current society. In the same way, this has led to the mass generation Jiménez et al., 2008). Different studies have also correlated this state
of other types of organic waste, such as those derived from the agri- with other factors such as enzymatic indicators or biodiversity profiles
cultural and agrifood industries, as well as the extraction of olive oil during composting (Jurado et al., 2014; López-González et al., 2015).
(Alburquerque et al., 2009; Wei et al., 2019). Currently, many strategies Therefore, at a legislative level, knowing the C/N ratio, moisture and
have been proposed for the management of organic waste (biowaste) in organic matter content of a certain compost is considered fundamental
a more sustainable manner. The composting process is considered one to determine if that product is appropriate from an agronomic point of
of the most attractive, in this sense, because it does not entail an en- view. However, (i) could it be thought that a compost will have a dif-
vironmental impact and does not imply a high cost. In fact, according to ferent lignocellulosic content depending on the raw material? (ii) or
the recommendation of the European Waste Law (EU Waste Law, that the biotransformation of organic waste occurs in the same way
2018), the composting process is positioned as the first and most ap- depending on the starting materials? In summary, is the composting
propriate alternative for the treatment and revaluation of biological process dependent or independent of the different raw materials?
waste. Raw materials used in composting are composed of both assimilable
Therefore, in order to control the quality of composting product, compounds and complex polymers. The most recalcitrant fractions,
based on its maturity, each country has established its own regulations such as lignocellulose, must be processed enzymatically before being
on the ideal characteristics that compost must possess to consider it used by microorganisms as carbon and nitrogen sources. This fraction,
with enough grade for soil application. In Spain, according to BOE A- whose biodegradation is important for the process of humification and
2017-14332, the final product of composting should meet the following stabilization at the end of the composting process, is difficult to

*
Corresponding author.
E-mail address: fsuarez@ual.es (F. Suárez-Estrella).

https://doi.org/10.1016/j.jenvman.2019.109312
Received 10 April 2019; Received in revised form 22 July 2019; Accepted 24 July 2019
0301-4797/ © 2019 Elsevier Ltd. All rights reserved.
M.J. Estrella-González, et al. Journal of Environmental Management 248 (2019) 109312

transform (Insam and de Bertoldi, 2007). However, although achieving 2.2. Basic parameters of control and monitoring of composting processes
an optimal degree of degradation is decisive to reach the appropriate
maturity for a quality compost, the enzymatic strategies and the hu- Several physicochemical parameters, based on those typically used
mification pathways could be very different in each process. for the control and monitoring of composting processes, were studied in
Enzymes produced by microorganisms participate in numerous this work.
biochemical transformations of organic matter occurred throughout the The moisture content (M) was determined by drying at 105 °C for
process (Mondini et al., 2004). Different studies have related the en- 24 h. The pH was analyzed in a 1:10 (w/v) water extract. Bulk density
zymatic biotransformations on lignocellulosic fractions with the com- (BD) was measured according to the US Composting Council USCC,
post stability degree (Ros et al., 2006; Jurado et al., 2015; Villar et al., 2001. Elemental carbon (EC) and nitrogen (N) were determined in solid
2016). In this sense, enzymatic evolution during the composting pro- samples by dry combustion at 950 °C using a LecoTruSpec C–N Ele-
cess could be considered an indicator of the material behavior and, mental Analyzer (Leco Co., St. Joseph, MI, USA). Organic matter con-
therefore, of the state of maturity (Castaldi et al., 2008). tent (OM) was assessed by determination of weight loss on ignition at
Since organic waste is constituted by a significant carbon fraction, 550 °C to a constant weight. Soluble organic carbon (SOC) was analyzed
enzymes related to carbon cycle are determinant during waste bio- in extracts obtained from fresh samples according to method described
transformation. β-glucosidase, amylase, cellulase and xylanase are by Hofman and Dusek (2003), and using a TOC-VCSN analyzer (Shi-
some of the most determinant enzymes in degradation of polymers and madzu, Japan). Extracts were prepared 1:4 (w:v) of fresh sample in
other saccharides during composting. Amylase is responsible for de- 0.5 M K2SO4, shaken at 200 rpm for 30 min, and filtered through filter
gradation of starch, resulting in glucose monomers. Xylanase is re- paper. The microbial biomass C (Cbio) was determined using the fumi-
sponsible for degradation of hemicellulose. Both, cellulase and β-glu- gation and extraction method according to Vance et al. (1987).
cosidase, participate in the degradation of cellobiose, and all of them
play an important role in the degradation of lignin (Jurado et al., 2014, 2.3. Lignocellulosic fractions
2015).
In order to elucidate the influence of raw materials on the biode- Cellulose (CEL), hemicellulose (HC) and lignin (LIG) fractions were
gradation dynamic of lignocellulosic fractions, and to determine the determined by a fiber analyzer ANKOM 200/220 (Ankom Technology,
exclusiveness of each composting process, stabilization processes of 5 Macedon, NY, USA). The methods applied in this case were those es-
different type of materials, including Vegetable Waste (VW), Urban tablished by ANKOM Technology for fiber analyses (http://www.ankom.
Solid Waste (USW), Agrifood Waste (AW), Sewage Sludges (SS) and com/procedures.aspx). Holocellulose was expressed as the sum of CEL
olive mill waste “Alpeorujo” (ALP), were analyzed in this work. To and HC values to calculate an additionally ratio, Lignin/Holocellulose.
verify this hypothesis, the following goals were set: (1) to analyse the Biodegradation ratios of HC, CEL and LIG were calculated according
conduct of physicochemical parameters related to the control and the following formula (Wang et al., 2011):
monitoring of the different composting processes, (2) to determine the
Rn (%) = [(mn mo )/ mn] × 100,
evolution of lignocellulose, hemicellulose, cellulose and lignin, as well
as different humification parameters, (3) to evaluate the effect of dif- where Rn is the degradation ratio (%) for nth sampling,
ferent enzymatic activities related to carbon fractions, and (4) to mo: initial content (mg g −1) of the lignocellulosic fraction (HC, CEL
stablish correlations between different groups of parameters on the or LIG),
basis of the diversity of the raw materials. This research will provide mn: content (mg g −1) of the lignocellulosic fraction (HC, CEL or
useful information about the way in which the processes behave, in- LIG) for nth sampling.
dependently of the starting material and the operational activities.
2.4. Enzymatic analysis
2. Material and methods
β-glucosidase, amylase, cellulase and xylanase activities were ana-
2.1. Sampling strategy and experimental set-up lyzed and the results were expressed as μmol product g−1 h−1 on a dry
weight basis.
Industrial processes carried out in 15 companies dedicated to the sta- The estimation of β-glucosidase activity was carried out following the
bilization of different organic materials were analyzed. Each one of them method described by Tabatabai (1982). This method is based on the col-
was chosen to obtain a collection of 270 samples (15 plants × 6 stages x 3 orimetric estimation of p-nitrophenol released by the hydrolysis of the p-
repetitions) made from different raw materials, and consequently to ob- nitrophenyl-β-D-glucopiranoside (25 mM pNPG in MUB-HCl buffer, pH6) at
serve the influence of starting waste on all the biotransformation pro- 37 °C for 1 h. Cellulase activity was determined by a modified method
cesses. The composting piles selected for this purpose were prepared described by Libmond and Savoie (1993), based on the colorimetric esti-
outside, in rows of 7–15 m in length and 3–5 m in width, and turned mation of the glucose released in the reaction occurred between the enzyme
periodically according to the periods of decrease of the internal tem- and the substrate (1% carboxy methyl cellulose in 50 mM sodium acetate
perature of the piles. Although each company works with mixtures of buffer, pH 5) at 37 °C for 2 h. The method described by He et al. (1993) was
different organic materials and in very variable proportions, the main applied for the detection of xylanase activity; this protocol was based on the
waste selected were as follow: Vegetable Waste (VW), Urban Solid Waste colorimetric estimation of the glucose released in the reaction occurred
(USW), Sewage Sludges (SS), Agrifood Waste (AW), and Olive oil mill between the enzyme and the substrate (1% xylan solution in 50 mM citrate-
wastewater “Alpeorujo” (ALP). Samples were collected at 6 critical periods phosphate buffer, pH 6.5) at 30 °C for 30 min. On the other hand, the
according to temperature evolution (including initial and final products): amylase activity was carried out following the method described by Mishra
Raw Materials (RM), Mesophilic phase (MES), Thermophilic phase et al. (1979), which consisted of a colorimetric estimation of the glucose
(THER), Cooling phase (COOL), Maturation phase (MAT) and Final released in the reaction occurred between the enzyme and the substrate
Product (FP). The temperature was periodically measured in situ with (1% soluble starch in 0.5 M phosphate buffer, pH 5.5) at 35 °C for 24 h.
long-handled PT100 probes (50 cm). During each stage, subsamples were
taken from nine different locations, covering several depths of each pile, 2.5. Humification parameters
by means of a soil sampling probe capable of reaching about 1–2 m depth.
Subsamples were mixed in equal amounts, in order to achieve a homo- Humic fractions were extracted and analyzed as described by
geneous and representative sample and, later, divided into three parts to Cavani et al. (2003). 2.0 g of sample were mixed with 100 mL of 0.1 M
work for analytical replicates. NaOH and 0.1 M Na4P2O7 x 10H2O and incubated for 48 h at 65 °C

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M.J. Estrella-González, et al. Journal of Environmental Management 248 (2019) 109312

Table 1
Evolution of moisture (M), pH, bulk density (BD), organic matter (OM), C/N ratio, soluble organic carbon (SOC) and biomass carbon (Cbio) throughout composting of
5 types of organic waste (USW: Urban Solid Waste; VW: Vegetable Waste; SS: Sewage Sludge; AW: Agrifood Waste; ALP: Alpeorujo). Composting phases: raw
materials (RM), mesophilic phase (MES), thermophilic phase (THER), cooling phase (COOL), maturation (MAT) and final product (FP). Nine repetitions have been
taken into account in each sampling. Values followed by same letters are not significantly different (P < 0.05) according to Fisher's LSD test.
Organic Waste Sampling M (%) pH BD (g/cm3) OM (%) C/N SOC (%) Cbio (%)

c a c d e c
USW RM 78.20 5.16 0.21 74.73 28.47 2.37 0.20c
MES 17.73a 7.84b 0.14a 66.26c 20.16d 1.23b 0.04a
THER 36.70b 8.43d 0.22cd 54.14a 14.03c 0.62a 0.12b
COOL 21.07a 8.12c 0.18b 57.60b 15.76b 0.51a 0.05a
MAT 13.05a 8.64d 0.25d 51.88a 12.09a 0.63a 0.05a
FP 11.31a 8.66e 0.22bc 53.91a 11.79a 0.61a 0.04a
VW RM 85.94f 8.10c 0.12ab 74.06e 12.63ab 0.49a 0.05ab
MES 77.05e 8.43d 0.10a 70.33d 14.18bc 0.52a 0.05ab
THER 46.97d 5.57a 0.16bc 69.35d 15.45c 1.41c 0.11d
COOL 53.42c 6.93b 0.18cd 63.20c 15.05c 1.36c 0.08c
MAT 43.92b 8.87e 0.19cd 58.85b 12.34a 0.58ab 0.06b
FP 41.05a 9.18e 0.22d 48.43a 11.73a 0.68b 0.03a
SS RM 81.87e 7.95b 0.28b 71.97e 9.29e 0.25c 0.06a
MES 67.09c 8.47d 0.19a 58.05d 7.32b 0.05a 0.10b
THER 73.68d 8.59de 0.23a 57.18d 8.42c 0.13b 0.10b
COOL 38.23b 8.62e 0.42c 49.40b 8.24a 0.15b 0.15c
MAT 35.74ab 8.27c 0.44c 53.38c 6.75a 0.22c 0.16c
FP 30.80a 7.72a 0.40c 47.19a 8.92d 0.27c 0.15c
AW RM 65.59b 6.67a 0.21a 62.51b 16.66b 0.13ab 0.14c
MES 66.60b 7.41b 0.20a 72.95c 19.16d 0.18bc 0.08b
THER 45.56b 8.44c 0.21a 66.96b 17.74c 0.26d 0.08b
COOL 33.16a 8.60d 0.27b 50.64a 12.51a 0.22cd 0.03a
MAT 35.15a 8.72e 0.30b 52.73a 12.69a 0.10a 0.03a
FP 30.45a 8.67de 0.29b 52.10a 11.83a 0.14ab 0.04a
ALP RM 62.48e 5.52a 0.18a 74.85c 19.02b 1.55c 0.05b
MES 57.36d 5.91ab 0.16a 73.52c 19.96b 1.08b 0.03ab
THER 51.17bc 6.15b 0.14a 75.60c 23.97c 1.40c 0.05ab
COOL 48.01b 9.17c 0.28b 60.07b 14.82a 0.65a 0.04ab
MAT 51.68c 9.28c 0.33c 55.90a 14.45a 0.46a 0.05b
FP 42.01a 9.46c 0.26b 60.23b 15.65a 0.73a 0.03a

under continuous stirring (120 rpm) in a thermostatic water bath. After Bioeconomy (EU Waste Law, 2018). With this idea it is intended that
extraction, the samples were centrifuged at 10000 g for 10 min and the materials do have a circular return to their origins. It means, to be
supernatants were filtered through a 0.8 μm filter (Standard MF-Milli- transformed into others less harmful to the environment and living beings.
pore Membrane, EMD Millipore Corporation, Billerica, MA, USA). This The regulations that entered into force establish very ambitious targets for
filtered solution constituted the total extractable carbon (TEC) that was recycling, setting several periods for the fulfillment of very strict objectives
fractioned additionally in humified (humic and fulvic acids) and not related to the reduction of residues, including organic waste. Specifically,
humified fractions. Total Organic Carbon (TOC), Total extractable before 2023, the collection and transformation of so-called biological
carbon (TEC), non-humified carbon (CNH), humic-like carbon (CHA) and wastes should be managed in a very controlled way. In this sense, without
fulvic-like carbon (CFA) were evaluated by means of carbon analysis in a doubt, composting is positioned as one of the most important tools for
the corresponding fractions (TOC-VCSN, Shimadzu, Japan). In addition, the future of biological waste management (EU Waste Law, 2018).
humification rate was calculated: HR = % ((CHA + CFA)/TOC) x100). Although the current European legislation on waste raises a series of
recommendations on the management of organic waste through com-
posting, each member state is governed by a series of specific regula-
2.6. Statistical analyses
tions. In the particular case of Spain, this is included in the Official State
Bulletin (BOE A-2017-14332 on fertilizer products). In above regula-
Data were subjected to a statistical analysis using Statgraphics
tions, reference is made to the basic characteristics that a compost must
Centurion XVI.I (StatPoint Technologies Inc., Virginia). Analysis of variance
have in order to be considered of quality, with special emphasis on
(ANOVA) and multiple comparison tests (Least Significant Difference, LSD)
parameters such as humidity (< 40%), minimum content in organic
were performed to compare the mean values of the analyzed variables and
matter (35–45%) or C/N ratio (< 15–20).
determine the influence of the different selected factors (p < 0.05).
The temperature inside the pile is one of the main factors to con-
Discriminant analyses were used to separate groups of independent vari-
sider during a composting process, since it directly affects the rate of
ables according to the stages of composting and the type of raw materials.
degradation of organic matter as well as determines the stability of the
Likewise, the relations between pairs of variables for each starting material
product. Stentiford (1996) suggested that temperatures above 55 °C
were analyzed using the Pearson correlation coefficient.
maximize the sanitation of materials, while temperatures between 45
and 55 °C improve biodegradation rates. According to the Environ-
3. Results and discussion mental Protection Agency (EPA, 2003), during the thermophilic phase
of a composting process, temperatures above 55 °C must be reached for
3.1. Monitoring of the control parameters during the composting processes 3 consecutive days. In this research work, 15 industrial-scale com-
on an industrial scale: moisture, pH, bulk density (BD), organic matter posting plants dedicated to the management of different types of bio-
(OM), C/N ratio, soluble organic carbon (SOC) and biomass carbon (Cbio) waste were selected. The average temperature in the thermophilic stage
for each installation met the minimum established by the EPA. The
The new legislation on waste is clearly oriented to the needs of the highest average temperature was reached in the VW piles (65 °C). In the
population in relation to the current concept of Circular Economy or

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M.J. Estrella-González, et al. Journal of Environmental Management 248 (2019) 109312

Fig. 1. Evolution (a, c, e, g, i) and biode-

gradation ratio (b, d, f, h, j) of cellulose CEL
(●), hemicellulose - HC ( ), lignin - LIG ( )
and lignin/holocellulose ratio - L/H (⊗)
throughout composting of 5 types of or-
ganic waste. Values are the mean of nine
replicates. LSD values are independently
shown for each parameter and composting
process: USW (LSDCELa = 0.9349;
LSDHCa = 1.3582; LSDLIGa = 0.2018; LSDL/
Ha = 0.0083, LSDCELb = 5.1583; LSDHCb =
9.3066, LSDLIGb = 3.0716), VW (LSDCELc =
0.7784; LSDHCc = 1.1319; LSDLIGc =
0.5046; LSDL/Hc = 0.0275; LSDCELd =
2.5893, LSDHCd = 9.9096, LSDLIGd =
3.9493), SS (LSDCELe = 0.9405; LSDHCe =
1.1049; LSDLIGe = 0.2886; LSDL/He =
0.0828; LSDCELf = 4.0721, LSDHCf =
8.3252, LSDLIGf = 2.9866), AW (LSDCELg =
1.0206; LSDHCg = 1.1029; LSDLIGg =
0.3964; LSDL/Hg = 0.0313; LSDCELh =
5.7033, LSDHCh = 6.9002, LSDLIGh =
3.0402) and ALP (LSDCELi = 0.6037;
LSDHCi = 0.9257; LSDLIGi = 0.3036; LSDL/
Hi = 0.0284; LSDCELj = 1.8968, LSDHCj =
5.7578, LSDLIGj = 4.3222).

rest of composting plants, the average temperature in the thermophilic Table 1 shows the values of the control parameters that were used to
phase ranged between 50 and 55 °C. All temperature data were pro- verify the composting processes evolution. Although the raw material
vided by the responsible companies in each case. The mean values of (RM) and the final product (FP) are not real phases of the composting
temperature as a function of the starting materials are provided in Table process, it is important to highlight the relevance of both stages to
S1 (see Supplementary material, Table S1). evaluate the optimal evolution of the composting process. In fact, in this

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M.J. Estrella-González, et al. Journal of Environmental Management 248 (2019) 109312

work, the bioxidative phase involves raw materials (RM), mesophilic the decomposition of this fraction in the materials that are being com-
(MES) and thermophilic (THER) stages, while the maturation phase posted. In any case, closely monitoring the changes that this fraction un-
refers to the pre-maturation (MAT), cooling (COOL) and final product dergoes during composting, can help to understand how the contribution
(FP) stages. With respect to the results shown in Table 1, in all cases, the of readily available compounds for growth and microbial activity occurs.
average moisture values were higher during the bioxidative phase of Adequate degradation of the lignocellulose fraction is crucial to achieve a
the composting process. The initial humidity of the piles was optimal in mature product, free of toxicity to be applied in agricultural soils.
the case of the facilities dedicated to AW and ALP, while this parameter In general terms, cellulose, hemicellulose and lignin decreased
was much higher than recommended in those companies dedicated to along the composting process, though this decrease was more notice-
USW, VW and SS. During the process, the moisture content contributes able in the case of the cellulosic fraction. The lignin/holocellulose (L/H)
to a better degradation of organic matter and maintenance of the ratio shows a tendency to increase at the end of the process, as a result
temperature during a longer period of time, because it favors the mi- of the transformation of lignocellulose (Fig. 1a, c, e, g, i). Exceptionally,
crobial activity (Silva et al., 2013). However, at the end of composting, in the case of the processes elaborated from plant wastes (VW), cellu-
the reduction in moisture content is a positive sign of decomposition lose remained practically constant in the intermediate stages, to then
and compost maturity (Ameen et al., 2016). In all the analyzed pro- decrease at the end of the process. Meanwhile, hemicellulose and lignin
cesses the humidity of the piles decreased over time to reach values decreased continuously. The L/H ratio showed a tendency to increase in
around 30–40% in the final products. Only in the case of products made the intermediate stages, although the final value was slightly lower than
from USW, moisture values were detected around 11%, which were that obtained in the initial stage (Fig. 1c). This fact could be due to the
considered excessively low (Table 1). accidental introduction of fresh material during the process.
Bulk density (BD) is an important factor to condition the initial The degradation degree of all the lignocellulosic fractions was very
mixtures in the composting processes. BD also influences the mechan- variable. Cellulose degradation reached values around 25–50% at the
ical properties of the compost, such as strength, porosity and ease of end of the processes made from USW, VW, AW and ALP (Fig. 1b, d, h, j)
compaction (Agnew and Leonard, 2003). In this work, the values of pH while it was higher than 80% in the case of SS processes (Fig. 1f).
and BD increased throughout the composting process (Table 1). The Hemicellulose degradation ranged between 20 and 80% approximately,
final pH values of all the compost oscillated around alkaline values. being minimal in the case of USW processes and maximum in the case
Singularly high were the pH values of the compost based on VW and of SS processes. Lignin, on the other hand, did not exceed 20% de-
ALP (López-González et al., 2015). gradation (Fig. 1b, f, h, j), excepting in the case of VW processes where
Regarding the percentage of organic matter (OM), it decreased lignin degradation almost reached 40% (Fig. 1d).
considerably in all the processes, which supports the fact that in all The starting material used in each process consisted of 12–15%
cases the biotransformation of the starting materials was being carried cellulose, and 4–8% hemicellulose and lignin. In general, the fraction
out. The initial content in OM ranged between 62 and 74%, detecting most affected by the composting process was cellulose, in contrast to
the highest loss of organic matter at the end of the USW (27%), VW lignin and hemicellulose, whose concentrations remained almost un-
(34%) and SS (34%) processes. The percentage of loss of organic matter changed compared to their content in raw materials. Other authors
for the other samples (AW and ALP) ranged between 16 and 19% observed this same effect in previous works (Jurado et al., 2014). In
(Table 1). Although in some cases the final OM values were somewhat some cases, the degradation of cellulose became more noticeable when
high, coinciding with what was obtained by other authors (Jurado a substantial amount of hemicellulose had been degraded. This fact may
et al., 2015; López-González et al., 2015). be due to the fact that the lignin-hemicellulose complex between the
At the beginning of composting, a C/N ratio around 25–30 is con- cellulose fibers decreases the available surface area, and prevents easy
sidered optimal; however, these initial values were only observed in the access to cellulose by the microorganisms of the process and its en-
case of processes made from USW. This fact confirms that, in many zymes (Komilis and Ham, 2003). According to other authors, however,
cases, the conditioning of the starting materials in the industrial scale hemicellulose is the fraction of lignocellulose that is easier to degrade
composting is not entirely adequate, due to the accidental mixing of and generally decomposes to a greater extent, in comparison with cel-
fresh materials with other partially biotransformed ones, which nega- lulose and lignin. Taking into account that hemicellulose is a poly-
tively could affect the balance between C and N within the piles. Only saccharide with a lower molecular weight than cellulose and lignin, Wei
in the case of USW samples, the decrease in this parameter was greater et al. (2019) stated that in general that is subject to a greater loss during
than 50% compared to the initial values. According to Guo et al. composting. These authors also stated that the high temperatures
(2012), the C/N ratio directly influences the maturity of the product reached during the thermophilic phase are important for the degrada-
obtained, affecting other parameters. For example, when in a com- tion of hemicellulose, while the degradation ratio of cellulose/lignin
posting process the initial C/N ratio is not adequate, it evolves un- increases from the cooling phase. In fact, Tuomela et al. (2000), pre-
expectedly and influences the content of SOC and Cbio, which may even viously reported that cellulose and hemicellulose degrading micro-
increase at the end of the process (Table 1). On the contrary, soluble organisms require high temperatures, even above 60 °C, while most
organic carbon is a fraction that constitutes a rapid source of carbon degradation of lignin generally occurs at lower temperatures (Tuomela
and energy for growth and microbial activity and, in suitable com- et al., 2000).
posting, it is expected to decrease throughout the process (Jurado et al., Therefore, the results corresponding to the degradation of the lig-
2014, 2015). Whilst, probably related to an incorrect C/N ratio at the nocellulosic fractions showed different profiles depending on each type
beginning, Cbio did not show significantly lower values at the end of the of raw material. This fact confirms that the transformation of polymeric
process in the case of the VW and SS processes (Table 1). carbon into soluble, simple, and available carbon compounds for the
microorganisms present in the piles is unique and depends on the ma-
3.2. Evolution of lignocellulosic fractions during the composting processes terials used. In spite of this, the global tendencies of biodegradation
on an industrial scale profiles were very similar in all the composting processes evaluated on
an industrial scale, especially with regard to cellulose.
The evolution of lignocellulosic fractions and the changes in the de-
gradation rates of lignocellulose during industrial-scale composting of 3.3. Evolution profiles of β-glucosidase, amylase, cellulase and xylanase
different wastes are shown in Fig. 1. The biodegradation of lignin during activities during the composting processes
the composting process generally occurs late and at very low rates
(Malherbe and Cloete, 2002). In addition, it acts as a protective factor for The biodegradation profiles of the lignocellulosic fractions de-
the degradation of cellulose and hemicellulose. Both aspects complicate scribed above depend absolutely on the activity of the microorganisms

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M.J. Estrella-González, et al. Journal of Environmental Management 248 (2019) 109312

Fig. 2. Dynamics of β-glucosidase, Amylase, Cellulase and Xylanase activities throughout composting made of USW (a-d), VW (e-h), SS (i-l), AW (m-p) and ALP (q-t).
Values are the mean of nine replicates. LSD values are indicated for each enzymatic activity and raw material (Fisher LSD test at p > 0.05).

present in the composting processes analyzed (Wei et al., 2019). This hemicellulose represented in Fig. 1. Although in all cases an increase in
dependence refers more specifically to the activity of a group of en- the biodegradation towards the end of the composting process was
zymes of microbial origin directly related to the degradation of this type perceived, a more pronounced degradation peak was detected in the
of molecules. Although culturable microbial communities have not cooling phase in the case of the processes made of VW and AW (Fig. 1g
been analyzed in this work, a group of enzymes highly involved in the and i). This could be due to the complexity of the lignocellulosic ma-
degradation of very complex molecules has been selected. Such is the terial or the entry of semi-processed or fresh (untreated) materials in
case of cellulose, hemicellulose and lignin, which can serve as bioin- the intermediate stages of the composting process.
dicators of the optimal evolution of the composting processes. In this With regard to the plants elaborated from Sewage Sludges
way, Fig. 2 shows the evolution of the activities of β-glucosidase, (Figs. 2i–1), a clear downward trend of all the enzymatic activities
amylase, cellulase and xylanase in the different composting processes analyzed was observed. The most pronounced decrease was detected in
analyzed, depending on each raw material and the different stages of the β-glucosidase and cellulase activities, whereas it was more gradual
composting. in the case of the amylase and xylanase activities. In the latter case, an
Fig. 2a–d shows the trends of these four enzymatic activities in increase in the activity was observed even during the thermophilic
composting processes made from USW. All enzymatic activities ana- stage. It should be noted that the evolution profiles of the enzymes
lyzed decreased as the composting processes progressed. Even so, the involved in the degradation of lignocellulosic fractions in Sewage
activity of the β-glucosidase showed a more oscillating behavior, de- Sludges plants coincide with the higher rates of biodegradation of these
tecting important peaks of activity during the cooling phase of the fractions at the end of the process (Fig. 1h).
process. In this type of waste, the levels of cellulase and xylanase ac- With respect to the enzymatic activities corresponding to the pro-
tivity at the beginning of the processes were maximum, probably due to cesses elaborated from “Alpeorujo”, Fig. 2q–t shows identical profiles of
the nature of the starting materials. β-glucosidase, amylase and cellulase activities. Those were significantly
Fig. 2e–h correspond to the evolution of enzymatic activity in greater during the bioxidative phase of the processes and gradually
composting processes made from Vegetable Waste (VW). In general decreased towards the end of them. However, the evolution of the xy-
terms, the enzymatic activity of the samples decreased progressively lanase activity (directly related to the degradation of hemicellulose)
throughout the composting process, although in a more discrete way in seemed to be controlled by the other three activities indicated above,
the case of the cellulase and xylanase activities, or even increased to- since it was oscillating throughout the process, showing two peaks of
wards the end of the maturation phase (Fig. 2h). These activity profiles maximum activity at the beginning and then in the cooling phase
were very similar to those observed in the case of the processes made (Fig. 2t).
from AW as main residues (Fig. 2m–p). It should be noted that the in- In view of the results obtained, it can be concluded that the selection
crease in the xylanase activity at the end of both processes (VW and of raw materials strongly influenced the enzymatic profiles of the dif-
AW) is directly related to the biodegradation rates of cellulose and ferent composting processes analyzed. In general terms, all enzymatic

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M.J. Estrella-González, et al. Journal of Environmental Management 248 (2019) 109312

activities tended to decrease at the end of the process (Fig. 2 a-p). Table 2
However, in some cases, β-glucosidase activity increased during the Change produced in humification parameters throughout composting of dif-
bioxidative phase, showing an oscillating profile, which could be due to ferent residues (USW: Urban Solid Waste, VW: Vegetable Waste, SS: Sewage
the continuous release of molecules more readily available for micro- Sludge, AW: Agroindustrial Waste and ALP: Alpeorujo). Nine repetitions have
been taken into account in each sampling. Values of elemental carbon (EC),
organisms (Fig. 2a, q). This fact coincided mainly with those processes
total organic carbon (TOC), non humic carbon (CNH), humic/fulvic carbon ratio
in which the initial cellulase activity was remarkably high (USW and
(CHA/CFA) and Humification Ratio (HR) followed by same letters are not sig-
ALP, Fig. 2c and s). In all processes, amylase showed a similar activity
nificantly different (p > 0.05) by Fisher's LSD test (RM: Raw Material, FP:
profile, so that it was more or less constant during the first stages of the Final Product).
processes and then progressively decreased during the maturation
Organic Waste Sampling EC (%) TOC (%) CNH (%) CHA/CFA HR
phase to practically undetectable values at the end of the composting
process (Fig. 2b, f, 2i, 2n, 2r). Although β-glucosidase and amylase USW RM 40.33b 43.45b 0.86a 1.41a 6.55a
activities showed the same trend in all processes, the highest values FP 27.99a 31.34a 0.83a 1.78a 5.69a
were observed in those made from Vegetable Waste (VW). Other au- VW RM 33.74b 43.06b 0.20a 0.45a 9.16a
thors have detected similar values of β-glucosidase in composting FP 24.51a 28.16a 0.62b 0.54a 14.45b
SS RM 35.45b 41.84b 1.08b 1.25a 6.05a
processes from urban solid waste (Zang et al., 2018). FP 23.17a 27.44a 0.47a 1.32a 9.27b
Cellulase and xylanase activity profiles were generally downwards, AW RM 35.52b 36.34b 0.67b 0.59a 7.14a
except in the case of the processes made from VW and AW, where those FP 30.54a 30.29a 0.52a 0.74a 12.69b
activities increased towards the end of the composting process (Fig. 2g, ALP RM 36.06b 43.51a 0.73a 1.40a 6.48ab
FP 31.75a 35.02b 0.50a 2.85a 4.40a
h, 2°, 2p). In both cases, the activity profiles coincide with what was
observed in Fig. 1, since it was in the VW and AW processes that a
sudden change in the biodegradation rates of the lignocellulosic frac-
than 30% over the initial value, while in others it did not exceed 14%.
tions was observed from the cooling phase onwards. In contrast to β-
Many authors have proposed the loss of carbon as a parameter that can
glucosidase, cellulase and xylanase activities were strongly influenced
serve as an indirect indicator of the degree of maturity of the compost
by the availability of the different lignocellulosic fractions, so their
(Jurado et al., 2015).
enzymatic profiles were highly variable. As described by Wei et al.
In general terms, non-humified carbon (CNH) values tended to de-
(2019), xylanolytic activity in Sewage Sludges (SS) and “Alpeorujo”
crease during all the composting processes analyzed, with the exception
(ALP) showed maximum activity in the thermophilic stage. However,
of the process elaborated from Vegetable Waste (Table 2). On the other
Amira et al. (2012), observed that xylanolytic and cellulolytic activity
hand, CHA/CFA ratio increased in the final phase, mainly and sig-
in Agrifood Waste increased at the end of the process.
nificantly in the case of the USW, AW and ALP processes. The positive
trend of the values of the CHA/CFA relationship could be explained by
3.4. Evolution of humification indicators during the composting process
the typical humification process that occurs during the maturation
phase of a composting process (Iglesias-Jiménez et al., 2008). CHA/CFA
During the maturation phase of a composting process, the stabili-
ratio has been proposed as an important indicator of compost stability,
zation and humification of the materials is carried out, obtaining a non-
being higher in mature compost (Azim et al., 2017). Compared to fulvic
toxic and humified product, which is very useful from the agronomic
acids, humic acids are derived mainly from a more mature fraction of
point of view. On the contrary, the immaturity of a compost could cause
organic material. CHA include compounds that increase the cation ex-
serious phytotoxicity problems as well as the depletion of the cultiva-
change capacity in soils (Mindari et al., 2014). They are related to soil
tion soils. Therefore, determining the maturity degree of a compost
fertility, crop yield and improve the C, N, P, K and S contents (Zhang
through the appropriate indicators could be of great interest to control
et al., 2017). Meanwhile, CFA are characterized by having a lower
the quality of the final product (Jurado et al., 2015).
molecular weight, a high level of oxidation and a lower amount of C, N
Traditionally, the quality of a compost has been determined through
and P, but they also improve the capacity of cation exchange in soil.
a group of physical-chemical parameters, such as temperature, CO2,
Therefore, the CHA/CFA ratio reflects the degree of polymerization in
humic/fulvic acid ratio, NH4–N/NO3–N ratio, C/N ratio, pH, microbial
materials of humic nature. According to Iglesias-Jiménez et al. (2008),
biomass, enzymatic activity, germination of seeds and concentration of
CHA/CFA values above 1 indicate that humification is occurring, while
heavy metals (Castaldi et al., 2008). However, due to the great diversity
values of CHA/CFA above 1.6 indicate that the product is already ma-
of raw materials and the different composting protocols, it is necessary
ture. Our data revealed that the best degree of humification was ob-
to establish more precise maturity indices (Martínez et al., 2016).
tained in the case of the processes elaborated from ALP.
Taking into account that the degree of maturation is more relevant
In addition to the CHA/CFA ratio, humification rate (HR) was cal-
at the end of the composting process, the humification parameters were
culated. During composting, thanks to a combination of chemical and
analyzed in this work only in the final stage (PF) of the different pro-
biological transformations, the total organic carbon content decreases,
cesses. However, the analysis of the humification parameters in the raw
while the relative content of humic compounds increases. According to
materials at the beginning of the composting processes was also in-
several authors, it has been demonstrated that the humification rate
cluded, to compare the results between the initial and final phase (RM-
(HR) is an effective indicator of the formation of humic substances
FP) in terms of maturity. Based on the above, the main parameters
during the composting process, being able to establish with precision
analyzed to determine the evolution of the humified fractions during
the moment of the complete stabilization (Tittarelli et al., 2002). Based
the selected composting processes are shown in Table 2 The initial and
on the results shown in Table 2, the humification rate (HR) was in-
final values of elemental and organic carbon (EC and TOC) as well as
creased in the case of the processes elaborated from VW, SS and AW. In
the proportion of non-humified substances (CNH), are shown in this
the case of the processes elaborated from “Alpeorujo” (ALP), contrary
table. However, the data related to the humic/fulvic ratio (CHA/CFA), as
to what was observed in the case of the CHA/CFA ratio, a decrease in the
well as the humification rate (HR), were parameters of greater interest
humification rate was detected. This could be due to the fact that the
when determining the humification dynamic of the processes.
HR values depend on the total organic carbon content of the samples
With respect to the different parameters related to humification, it
and, probably, there could have been an entry of fresh material towards
should be noted that very different trends were observed in terms of
the end of the process, which stopped the mineralization process.
raw materials (Table 2). As expected, elemental and organic carbon (EC
Results described for all the composting processes analyzed, agree
and TOC) values decreased in all composting processes. In some cases
that during the degradation of different materials, the amount of non-
(USW, VW and SS), the decrease meant a total carbon reduction of more

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M.J. Estrella-González, et al. Journal of Environmental Management 248 (2019) 109312

humified compounds (CNH) extracted in alkaline conditions was sig- analyzed, Fig. 3c shows the existence of two functions that explained
nificantly reduced over time, except in the case of the processes ela- more than 94% of the variability of the data. Thus, surprisingly, the
borated from vegetable remains (Table 2). During the composting results were grouped into 3 data sets, one of them corresponding to the
process, the release of compounds that are incorporated into the hu- initial phase of the processes (RM = Raw Materials), a second group
mification routes takes place through numerous reactions of re- formed by the data related to the bioxidative phase (Phase
organization and molecular condensation (Alburquerque et al., 2009). I = MES + THER) and a third group including the data corresponding
This fact leads to the formation of more humified compounds. In this to the stabilization and humification phase (Phase II =
sense, it is known that lignin plays an essential role in the processes of COOL + MAT + FP). Therefore, notwithstanding the heterogeneity of
humification of lignocellulosic residues, since the partial degradation of the data in relation to the starting materials and working conditions of
this complex molecule can give rise to the basic structure of humic each company, the data are suitably grouped from an evolutionary
substances, as well as to the liberation of aromatic and phenolic by- point of view, establishing continuity and logical limits between the
products (Alburquerque et al., 2009; Jurado et al., 2015). All these three main stages of the process: Initial phase (RM), Bioxidative phase
phenomena contribute to increase the content of humified substances (Phase I) and Maturation phase (Phase II). The above statement may be
during the maturation phase of the composting process. However, the irrelevant when working with small-scale or semi-heavy composting.
type of humified substances (humic or fulvic acid) and the generation However, the control of the different thermal phases of the process is
speed of them can vary greatly depending on the starting materials. crucial when working on composting on an industrial scale, since the
accidental (or voluntary) mixing of raw materials with others that are
3.5. Establishment of the differential profiles based on the diverse raw already biotransformed or that are in an intermediate phase of trans-
materials: discriminant analysis and pearson correlation formation could happen.
In order to analyse the composting profiles from each kind of ma-
On the basis of the results described and above discussed, it is al- terial, five discriminant and Pearson correlation analyses were per-
most evident that different physical-chemical and enzymatic scenarios formed for each process independently scrutinized (Fig. 4). Fig. 4 a-e
can be found depending on the composted materials or even on each showed the profiles observed for each group of material, depending on
process analyzed. Numerous recent scientific studies highlight the im- the succession of thermal phases. In all cases, discriminant function
portant relationship between the enzymatic activities involved in the explained near or even more than 90% of the variability results. Each
biodegradation processes of the lignocellulosic fractions and the evo- profile was apparently different from the rest, which supported that
lution of the humification markers during the composting process each process is exclusive. Data were located in three main groups: RM,
(Awasthi et al., 2018; Song et al., 2018; Gao et al., 2019). However, the Phase I (MES + THER) and Phase II (COOL + MAT + FP). In general,
verification of the actual influence of the raw material as a starting bioxidative (Phase I) and stabilization-humification phases (Phase II)
point to obtain compost in different pathways was one of the most appeared well separated in all the cases (Fig. 4 a-e). However, only in
important challenges addressed in this work. the composting process performed from AW (Agrifood Waste), a logical
Fig. 3 shows the discriminant analysis that help to understand how succession among COOL, MAT and FP was observed (Fig. 4d). On the
distinctive or similar the composting processes are depending on the contrary, during the processes performed from USW and SS, the MAT
starting materials (Fig. 3a), the final products obtained (Fig. 3b) and the and FP stages appeared very closed between them (Fig. 4b–c), while
thermal phases of the processes (Fig. 3c). COOL stage appeared very near of FP in the case of composting pro-
The discriminant analysis shown in Fig. 3a revealed two dis- cesses made from VW and ALP (Fig. 4a, e).
criminant functions responsible for more than 80% of the variability To establish significant links between the parameters analyzed in
observed among the data. In view of what is observed in Fig. 3a, it is this study (lignocellulosic-humic fractions and enzymatic profiles),
clear that data related to the lignocellulosic fractions, enzymatic ac- Pearson correlation maps were made for each type of material
tivities and humification process are grouped separately on the basis of (Fig. 4f–j). Many surprising relationships were detected, but different in
the raw materials. In this Figure, the set of data corresponding to the each case. In the case of the VW and USW composting processes, there
compost elaborated from Vegetable Waste (VW) stands out as being the were no significant correlations between the humification parameters
most distant group in relation to the rest (Fig. 3a). When the same and the rest of the variables (Fig. 4g). However, only in the case of
analysis was exclusively performed bearing in mind the data set cor- compost samples from plant remains was a close negative relationship
responding to the final products (FP), two discriminant functions in- detected between the hemicellulose content and the CHA/CFA ratio
fluenced the variability of the data by more than 85% (Fig. 3b). Fig. 3b (Fig. 4f). Something similar happened in the case of the samples from
supports the diversity of compost due to different raw materials, since 5 ALP, although in this case the CHA/CFA ratio was negatively correlated
significantly different groups were stablished based on that criterium. with the cellulose fraction and the xylanase activity (Fig. 4j). On the
While products (compost) made from AW, SS and VW were more si- contrary, the Pearson correlation profiles were very different in the case
milar in terms of biological activity as well as content in lignocellulosic of samples made from SS and AW, since the humification rate (HR)
and humic fractions, compost from ALP and USW were very different significantly correlated with the different enzymatic activities con-
from the rest and from each other (Fig. 3b). Despite the remarkable cerning the biodegradation of the lignocelullosic fractions (Fig. 4h and
differences detected among compost made from different organic i). Pearson correlation map between lignocellulosic and enzymatic
waste, all the analyzed samples complied with the basic monitoring parameters was remarkable in the case of samples made from SS, since
parameters (Moisture, Organic Matter and C/N Ratio) indicated at the positive links were detected among the group integrated by β-glucosi-
legislative level (BOE A-2017-14332). In this sense, humidity data re- dase-amylase-cellulase activities (Fig. 4h). A more discrete correlation
lative to the compost samples ranged approximately between 30 and was stablished in the case of samples made from AW and ALP (Fig. 4i
40%, excepting the final products made from USW, where a very low and j).
humidity was detected at the end of the process (Fig. 3d). The organic In view of the results shown in the correlation maps (Fig. 4f–j), it is
matter content was greater than 35–45% in all the final products ana- obvious that the USW-based composting processes did not develop or-
lyzed but this parameter was significantly higher in the case of samples thodoxly, since during a biotransformation process of this nature, it is
from ALP. Both circumstances could be involved in the differences expected that some type of relationship will be established between the
observed in the compost ALP and USW with respect to the rest of degradation of the lignocellulosic fractions, the degree of humification
products from different origins (Fig. 3b). and the enzymatic activity (Fig. 4f). This connection did not occur in
On the other hand, in order to establish the global differences or this case.
similarities between the different phases of the 15 composting processes Some of the enzymatic activities showed homogeneous and

8
M.J. Estrella-González, et al. Journal of Environmental Management 248 (2019) 109312

Fig. 3. Discriminant analysis based on composting of different raw materials, taking into account the complete processes (a), the final products (b) and the dis-
tribution of principal thermal phases (RM = Raw Material; Phase I = MES+THER; Phase II: COOL+MAT+FP) (c). Range (minimum and maximum values)
corresponding to moisture, organic matter and C/N ratio for each type of residues are shown compared with those included in the Spanish regulation as the main
parameters of compost quality (d).

independent profiles of the type of raw material (section 3.2.). Such is have tried to select the most appropriate waste mixtures and the best
the case of β-glucosidase and amylase activities, wich established close indicators to control the start-up of the process (Ghinea et al., 2019). In
links with each other and with other related enzymatic parameters. this sense, some of the most interesting physical-chemical parameters
Surprisingly, the correlations did not always occur in the same direc- used for this purpose are pH, electrical conductivity, humidity, ash,
tion. In the case of plants dedicated to Sewage Sludges composting content of N and C, C/N ratio, total organic carbon or the presence of
(Fig. 4h), Pearson P values were positive and very close to 1 in the β- heavy metals. Other authors have tried to establish the biodegradable
glucosidase-amylase-cellulase-xylanase block. This correlation did not character of the starting materials in order to optimize the bio-
happen in the case of plants dedicated to AW composting, where very transformation process as much as possible (Gutiérrez et al., 2017).
strong and negative correlations were established between cellulase However, the results described here have revealed the exclusive nature
activity and the β-glucosidase-amylase pair (Fig. 4i). A totally opposite of composting processes on an industrial scale. This exclusivity is fun-
effect to the previous one was observed in the case of the ALP samples damentally based on the different raw materials and operating condi-
(Fig. 4j). tions. Therefore, similar products could have been obtained thanks to
Although the relationships established between many of the ana- different enzymatic and humification profiles. In spite of this, in all
lyzed parameters were variable depending on the raw materials, there cases, the tendencies of some enzymatic indicators such as β-glucosi-
is no doubt that some of the enzymatic activities, such as β-glucosidase dase and amylase were very homogeneous.
and amylase, showed similar trends during the composting process
(section 3.3). This fact makes them very useful bioindicators to evaluate
4. Conclusions
the correct or incorrect evolution of the composting processes in-
dependently of the starting materials. Something similar happened in
Although different compost could be similar in relation to their
the case of the humification rate (HR) since, although in general it
agronomic quality, the enzymatic strategies and the humification
showed a logical tendency to increase toward the end of the process
pathways could be very different in each process. The results derived
(Table 2), the Pearson values observed were highly variable because of
from this work admit that each composting process is unique and runs
the type of materials. In the case of the SS samples (Fig. 4h), HR showed
independently, even showing similar trends.
a negative correlation with the β-glucosidase-amylase-xylanase
Parameters such as β-glucosidase, amylase, lignin/holocellulose
package, whereas in the case of AW plants, the HR correlated negatively
ratio or the humification degree could act as important indicators of the
with the β-glucosidase-amylase pair, but in a positive way with the
process. This fact confirms the efficiency and versatility of microbial
cellulase-xylanase block. Despite the different established links, the
enzymes and their role as transforming agents of organic matter, even
control of the evolution of the humification rate (HR), in combination
when the heterogeneity of materials and operating conditions on in-
with other enzymatic parameters, could be useful to determine the
dustrial scale are very diverse.
correct or incorrect evolution of the composting processes.
Taking into account the initial considerations regarding the singu-
To establish the most suitable recipe for composting, some authors
larity of each composting process, it seems evident that there are not

9
M.J. Estrella-González, et al. Journal of Environmental Management 248 (2019) 109312

Fig. 4. Comparative analysis of the different composting processes according to the starting materials (USW: Urban Solid Waste; VW: Vegetable Waste; SS: Sewage
Sludges; AW: Agrifood Waste, ALP: Alpeorujo). Discriminant analysis according to the distribution of the main thermal phases (a-e) and Pearson tables showing
positive (light grey) or negative (dark grey) correlations between the different analyzed parameters (f-j). HA/FA: CHA/CFA; HR: Humification Rate; CEL: Cellulose;
HEM: Hemicellulose; LIG: Lignin; β-glucosidase, Amylase, Cellulase and Xylanase.
10
M.J. Estrella-González, et al. Journal of Environmental Management 248 (2019) 109312

two equal composting processes. This can be directly related to the xylan fractions. Enzym. Microb. Technol. 15, 13–18. https://doi.org/10.1016/0141-
starting materials, the physical-chemical and enzymatic profiles that in 0229(93)90110-N.
Hofman, J., Dusek, L., 2003. Biochemical analysis of soil organic matter and microbial
each process are outlined. This fact can be considered, therefore, as the biomass composition - a pilot study. Eur. J. Soil Biol. 39, 217–224. https://doi.org/
main impact derived from this work. There is no recipe or a fixed 10.1016/j.ejsobi.2003.08.002.
protocol for the practice of composting, but different starting situations Iglesias-Jiménez, E., Barral, M.T., Marhuenda, F.C., 2008. Indicadores de la estabilidad y
madurez del compost. In: Moreno, J., Moral, R. (Eds.), Compostaje. Mundi- Prensa,
must be control from the beginning. Madrid, Spain, pp. 243–283.
Although different composting processes follow different paths, it is Insam, H., de Bertoldi, M., 2007. Microbiology of the composting process. In: Diaz, L.F.,
possible to reach to final products that comply with the legally estab- de Bertoldi, M., Bidlingmaier, W., Stentiford, E. (Eds.), Compost Science and
Technology. Elsevier, Amsterdam, The Netherlands, pp. 26–48.
lished quality criteria. So, in this work, final products that complied Jurado, M.M., Suárez-Estrella, F., Vargas-García, M.C., López, M.J., López-González, J.A.,
with the legal recommendations (minimum content in organic matter, Moreno, J., 2014. Evolution of enzymatic activities and carbon fractions throughout
humidity and C/N ratio) obtained. composting of plant waste. J. Environ. Manag. 133, 355–364. https://doi.org/10.
1016/j.jenvman.2013.12.020.
On the other hand, we must highlight the role of certain enzymatic
Jurado, M.M., Suárez-Estrella, F., López, M.J., Vargas-García, M.C., López-González, J.A.,
activities, such as β-glucosidase and amylase, as well as the ratio lignin/ Moreno, J., 2015. Enhanced turnover of organic matter fractions by microbial sti-
holocellulose, as bioindicators to monitor the composting process. The mulation during lignocellulosic waste composting. Bioresour. Technol. 186, 15–24.
profiles observed in this sense followed a characteristic and in- https://doi.org/10.1016/j.biortech.2015.03.059.
Komilis, D.P., Ham, R.K., 2003. The effect of lignin and sugars to the aerobic decom-
dependent pattern of the starting materials. position of solid wastes. Waste Manag. 23 (5), 419–423. https://doi.org/10.1016/
S0956-053X(03)00062-X.
Acknowledgements Libmond, S., Savoie, J., 1993. Degradation of wheat straw by a microbial commu-
nitystimulation by a polysaccharidase complex. Appl. Microbiol. Biotechnol. 40,
567–574. https://doi.org/10.1007/BF00175749.
This work has been funded by the Ministry of Economy and López-González, J.A., Vargas-García, M.C., López, M.J., Suárez-Estrella, F., Jurado, M.M.,
Competitiveness through project AGL2015-64512R. Moreno, J., 2015. Biodiversity and succession of mycobiota associated to agricultural
lignocellulosic waste-based composting. Bioresour. Technol. 187, 305–313. https://
doi.org/10.1016/j.biortech.2015.03.124.
Appendix A. Supplementary data Malherbe, S., Cloete, T.E., 2002. Lignocellulose biodegradation: fundamentals and ap-
plications. Rev. Environ. Sci. Biotechnol. 1 (2), 105–114. https://doi.org/10.1023/
A:102085891.
Supplementary data to this article can be found online at https:// Martínez, M., Ortega, R., Janssens, M., Angulo, J., Fincheira, P., 2016. Selection of ma-
doi.org/10.1016/j.jenvman.2019.109312. turity indices for compost derived from grape pomace. J. Soil Sci. Plant Nutr. 16 (2),
262–267. https://doi.org/10.4067/S0718-95162016005000021.
Mindari, W., Aini, N., Kusuma, Z., Syekhfani, S., 2014. Effects of humic acid-based cation
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