CN117678727B

CN117678727B - Use of lactobacillus fermentation in the preparation of a flavoring

Info

Publication number: CN117678727B
Application number: CN202311725265.3A
Authority: CN
Inventors: 刘晓勇; 金相武
Original assignee: Shandong Haizhibao Seafood Co ltd
Current assignee: Shandong Haizhibao Seafood Co ltd
Priority date: 2023-12-14
Filing date: 2023-12-14
Publication date: 2024-07-16
Anticipated expiration: 2043-12-14
Also published as: CN117678727A

Abstract

The invention discloses an application of lactobacillus to fermentation of a stannum algae processing byproduct in preparing a seasoning and the seasoning, wherein the seasoning comprises the lactobacillus-fermented stannum algae processing byproduct, solar salt, nucleic acid, glucose, red pepper powder, onion powder, ginger powder, garlic powder and bean paste powder. The flavoring is prepared by using the tannery algae processing byproducts fermented by lactic acid bacteria, so that the peculiar smell of the tannery algae processing byproducts is removed, the production cost can be saved, the profit of a company is increased, and the environmental problem is solved.

Description

Use of lactobacillus fermentation in the preparation of a flavoring

Technical Field

The invention relates to the technical field of seasonings, in particular to application of lactobacillus fermentation in preparing seasonings.

Background

Algae have long been used as a valuable food resource for humans. Seaweed is rich in vitamins, fibers and non-digestible viscous polysaccharides not present in terrestrial plant sources (1). Because seaweed has various biological activities such as anticoagulation, prevention of oxidative cell damage, antioxidation and immunomodulation, seaweed becomes an important functional and nutritional resource for developing and producing useful therapeutic agents and nutrient-rich food materials (2, 3, 4). Ceylon algae (Gelidium amansii, agar) is a red algae, mainly used for the production of agar which is insoluble in cold water but soluble in boiling water. The 1.5% agar solution was clear and when cooled to 34-43 ℃, the agar formed a firm gel. Agar is thus a mixture of polysaccharides, the basic monomer of which is galactose. Although the by-product of the tannery algae is discarded or used only as fertilizer, it appears to retain a variety of nutritional and functional components including polysaccharides and proteins. If the Ceylon algae byproducts are developed into more value-added raw materials, the environmental problem can be solved, and additional economic benefits can be obtained.

The seaweed (5) is processed using thermal extraction, enzymatic hydrolysis, acid extraction or alkali extraction. However, these processes often require expensive equipment or may degrade the functional compounds. Thus, there is a need in the art for a method of improving CMBP quality characteristics of seaweed to remove off-flavors from seaweed that is simple to operate, does not destroy the functional compounds, and utilizes CMBP to prepare functional food supplements or more value-added raw materials.

Disclosure of Invention

As previously mentioned, there is a need in the art for a method of improving CMBP quality characteristics to remove the off-flavors of seaweed and thereby prepare functional food supplements and the like. The inventors obtained a seasoning with sensory scores similar to other commercial products using CMBP of lactic acid bacteria fermentation as a raw material.

In a first aspect, the invention provides the use of lactic acid bacteria for fermenting a tannery algae processing byproduct (celyon moss processing by-product, CMBP) for the preparation of a flavoring.

In a second aspect, the present invention provides a seasoning comprising lactobacillus-fermented tannery algae processing byproducts.

The beneficial effects of the invention include:

(1) Removing the peculiar smell of the seaweed by utilizing a fermentation technology, and preparing the Ceylon seaweed processing byproducts with high protein and high carbohydrate content and low fat content into more value-added products;

(2) Saving production cost, increasing company profits and solving environmental problems.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings in the embodiments will be briefly described below.

FIG. 1 shows the total viable cell count (A) and the lactic acid bacteria count (B) of CMBP fermented by lactic acid bacteria at different fermentation periods.

FIG. 2 shows the pH (A), TA (B) and reducing sugar content (C) of CMBP fermented by lactic acid bacteria at different fermentation periods.

FIG. 3 shows the amino nitrogen content of CMBP fermented by lactic acid bacteria during different fermentation periods.

FIG. 4 shows a two-dimensional (2D) principal component analysis of CMBP extracts fermented for 48 h.

Detailed Description

Hereinafter, the present invention will be described in detail with reference to the accompanying drawings. It is to be understood that the following description is intended to illustrate the invention by way of example only, and is not intended to limit the scope of the invention as defined by the appended claims. And, it is understood by those skilled in the art that modifications may be made to the technical scheme of the present invention without departing from the spirit and gist of the present invention. The technical means used in the examples are conventional means well known to those skilled in the art unless otherwise indicated.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the subject matter described herein belongs. Before describing the present invention in detail, the following definitions are provided to better understand the present invention.

Where a range of values is provided, such as a range of concentrations, a range of percentages, or a range of ratios, it is to be understood that each intervening value, to the tenth of the unit of the lower limit, between the upper and lower limit of the range, and any other stated or intervening value in that stated range, is encompassed within the subject matter unless the context clearly dictates otherwise. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and such embodiments are also included in the subject matter, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the subject matter.

In the context of the present invention, many embodiments use the expression "comprising", "including" or "consisting essentially/mainly of … …". The terms "comprises," "comprising," or "consists essentially of … …" are generally understood to be open ended terms that include not only the individual elements, components, assemblies, method steps, etc., specifically listed thereafter, but also other elements, components, assemblies, method steps. In addition, the expression "comprising," "including," or "consisting essentially of … …" is also to be understood in this document as a closed-form expression, in certain instances, to mean that only the elements, components, assemblies, method steps specifically listed thereafter are included, and that no other elements, components, assemblies, method steps are included. At this time, the expression is equivalent to the expression "consisting of … …".

For a better understanding of the present teachings and without limiting the scope of the present teachings, all numbers expressing quantities, percentages or proportions used in the specification and claims, and other numerical values, are to be understood as being modified in all instances by the term "about" unless otherwise indicated. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained. At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

Herein, the term "lactic acid bacteria (LACTIC ACID bacteria, LAB)" refers to a group of gram-positive, catalase-negative, non-motile, microaerophilic or anaerobic bacteria that ferment sugars and produce acids, including lactic acid (as the primary acid produced), acetic acid, formic acid and propionic acid. Lactic acid bacteria most useful industrially include, but are not limited to, lactobacillus species (Lactobacillus spp.), lactococcus (Lactococcus spp.), streptococcus (Streptococcus spp.), leuconostoc (Leuconostoc spp.), pediococcus (Pediococcus spp.), brevibacterium (Brevibacterium spp.), enterococcus (Enterococcus spp.), and Propionibacterium (Propionibacterium spp.). In addition, lactic acid producing bacteria belonging to the strict anaerobe group, bifidobacteria (Bifidobacteria), i.e. Bifidobacterium spp, are often used alone or in combination with lactic acid bacteria as food starter agents, typically included in the lactic acid bacteria group. Even certain bacteria of the genus Staphylococcus (Staphylococcus) (e.g., staphylococcus botulinum (s. Carnosus), staphylococcus equi (s. Equum), staphylococcus murine (s. Sciurii), staphylococcus calf (s. Vitulinus) and Staphylococcus xylosus (s. Xylosus)) are known as LABs (Seifer and Mogensen (2002)).

As used herein, the term "Ceylon algae (ceylon moss)" is a red algae, namely Gelidium amansii (Gelidium amansii), which can be used to extract agar. There are agar dishes in each big sea area of China. The agar grows on the reefs in the shallow water, and the color is mauve, brownish red, faint yellow and the like, and the agar is also called sarcandra glabra because of the shape of the agar as coral. In addition, agar is called as herba Origani or frozen vegetable in Bohai sea coastal region, and is called as "agar" or "red silk" in Fujian region. The agar is transparent and has a plurality of irregular branches like a small tree. The agar is also known as carrageen by people in the form of carrageen. The Shennong Ben Cao Jing records agar: sweet and salty taste, severe cold, slippery and nontoxic, and mainly removes floating heat of upper energizer and deficiency-cold in lower part. The agar-agar has the effects of clearing lung-heat, resolving phlegm, cooling blood, stopping bleeding, nourishing yin, reducing pathogenic fire and the like, and is mainly used for treating symptoms such as cough with phlegm due to lung heat, constipation due to intestinal dryness and the like. The agar is rich in minerals, polysaccharide substances and multiple vitamins, and has high edible value.

As used herein, the term "agar" refers to a hydrocolloid extracted from certain seaweed of the class Rhodophyceae. It is insoluble in cold water but soluble in boiling water. The 1.5% agar solution was clear and when cooled to 34-43 ℃, the agar formed a firm gel. Agar is a mixture of polysaccharides, the basic monomer of which is galactose.

Herein, the term "electronic Nose" refers to any device designed to measure the gaseous chemical content of a sample of air or other gas, and includes devices that identify a particular component of an odor and analyze its chemical composition to identify it. The electronic nose may be composed of a mechanism for chemical detection (such as an array of electronic sensors) and a mechanism for pattern recognition (such as a neural network). The descriptors of the smell can then be extracted from the sensor itself or from pattern recognition.

In this context, amino acids are indicated by single-letter and three-letter abbreviations well known in the art. For example, glycine is represented by Gly or G, alanine is represented by Ala or A, valine is represented by Val or V, leucine is represented by Leu or L, isoleucine is represented by Ile or I, proline is represented by Pro or P, phenylalanine is represented by Phe or F, tyrosine is represented by Tyr or Y, tryptophan is represented by Trp or W, serine is represented by Ser or S, threonine is represented by Thr or T, cysteine is represented by Cys or C, methionine is represented by Met or M, asparagine is represented by Asn or N, glutamine is represented by Gln or Q, aspartic acid is represented by Asp or D, glutamic acid is represented by Glu or E, lysine is represented by Lys or K, arginine is represented by Arg or R, histidine is represented by His or H. Herein, the term "sweet amino acid (sweet amino acids)" refers to threonine, proline, serine, glycine, alanine, and lysine; the term "salty amino acid (savory amino acids)" refers to aspartic acid, glutamic acid, and cysteine; the term "bitter amino acid (bitter amino acids)" refers to methionine, isoleucine and leucine.

Studies have reported that microbial fermentation may be an alternative method of increasing the extractability of biologically active compounds. Lactic acid bacteria (LACTIC ACID bacteria, LAB) have been widely used as microbial food supplements with beneficial effects in humans, such as reducing lactose intolerance, lowering blood cholesterol, enhancing immune responses, preventing cancer. Lactic acid bacteria also produce new substances through bioconversion, lactic acid production, pickled cabbage products, plant defense stimulators, and antioxidant enhancement and anticoagulant activity. However, there are few reports on the use of seaweed biomass, especially as natural flavoring.

As previously mentioned, the Ceylon algae processing by-product (CMBP) has high protein and high carbohydrate content, low fat content, and the present invention aims to provide a method for improving CMBP quality characteristics by fermenting CMBP with lactic acid bacteria to remove the off-flavor of algae, and obtain a seasoning with similar sensory scores to other commercial products.

In a first aspect, the present invention provides the use of lactic acid bacteria to ferment a tannery processing byproduct in the preparation of a flavoring. In one embodiment, the lactic acid bacteria may be lactobacillus bulgaricus (Lactobacillus bulgaricus, LB), lactobacillus plantarum (Lactobacillus plantarum, LP), lactococcus lactis (Lactococcus lactis, LL) or streptococcus thermophilus (Streptococcus thermophillus, ST). In a preferred embodiment, the lactic acid bacteria are lactobacillus bulgaricus. It will be appreciated that other suitable lactic acid bacteria are also suitable for use in the present invention. In a further embodiment, the time to ferment the tannery algae processing byproduct using the lactic acid bacteria may be 48 hours. It is to be understood that other suitable times are also suitable for use with the present invention.

In a second aspect, the present invention provides a seasoning comprising lactobacillus-fermented tannery algae processing byproducts. In one embodiment, the lactic acid bacteria may be lactobacillus bulgaricus (Lactobacillus bulgaricus, LB), lactobacillus plantarum (Lactobacillus plantarum, LP), lactococcus lactis (Lactococcus lactis, LL) or streptococcus thermophilus (Streptococcus thermophillus, ST). In a preferred embodiment, the lactic acid bacteria are lactobacillus bulgaricus. It will be appreciated that other suitable lactic acid bacteria are also suitable for use in the present invention. In a further embodiment, the time to ferment the tannery algae processing byproduct using the lactic acid bacteria may be 48 hours. It is to be understood that other suitable times are also suitable for use with the present invention.

In one embodiment, the flavoring further comprises sun-dried salt, nucleic acid, glucose, red pepper powder, onion powder, ginger powder, garlic powder, soy sauce powder. In a further embodiment, the lactobacillus-fermented tannery algae processing byproduct, solar salt, nucleic acid, glucose, red pepper powder, onion powder, ginger powder, garlic powder, soybean paste powder may have a mass ratio of 60:40:10:20:10:2:1:4:6, but is not limited thereto. In a specific embodiment, the flavoring of the present invention comprises the following ingredients: 15g of lactobacillus-fermented tannery algae processing byproducts, 10g of sun-dried salt, 2.5g of nucleic acid, 5g of glucose, 2.5g of red pepper powder, 0.5g of onion powder, 0.25g of ginger powder, 1g of garlic powder and 1.5g of bean paste powder.

Examples

The conception and the technical effects produced by the present invention will be clearly and completely described in conjunction with the embodiments below to fully understand the objects, features and effects of the present invention. It is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments, and that other embodiments obtained by those skilled in the art without inventive effort are within the scope of the present invention based on the embodiments of the present invention.

Example 1

In this example, a Control group (Control, CMBP without lactobacillus fermentation) was set up and compared with four experimental groups (CMBP with four lactobacillus fermentations added) to compare the volatile compound profile, amino acid composition and sensory evaluation between the different groups.

The materials and methods used in this example are as follows:

The material Ceylon algae byproduct powder was obtained from Korea Min-Sok Food (Donghae, korea) and stored at-25℃for use. Trinitrobenzenesulfonic acid (TNBS), naOH, 3, 5-dinitrosalicylic acid (DNS) reagent, trichloroacetic acid (TCA), K ₂CO₃ and H ₂SO₄ were purchased from Sigma-ALDRICH CHEMICAL co. (St.Louis, MO, USA). Man Rogosa Sharpe (MRS) and Brain Heart Infusion (BHI) media were purchased from Merck (DARMATADT, germany). All other reagents used were of analytical grade.

Approximation composition analysis the approximation composition (including moisture, crude protein, fat, ash and carbohydrates) of CMBP was determined according to the official analytical chemist's society (AOAC) (6).

Preparation of starter culture pure cultures of lactobacillus bulgaricus ATCC 7995 (LB), lactobacillus plantarum ATCC 8014 (LP), lactococcus lactis ATCC 11454 (LL) and streptococcus thermophilus ATCC 14485 (ST) were obtained from korean microorganism culture center (KCCM, head, korea). LB, LP and ST were cultured using MRS culture at 37℃and LL was cultured using BHI culture at 37 ℃.

Preparation of fermentation CMBP in a 1L Elenmeyer flask, 15g of CMPB powder was homogenized with 300mL of distilled water, heated at 80℃for 30min, and then cooled to room temperature. After filtration using an 80mm sieve, the CMBP% LAB extract was inoculated with. Subsequently, CMBP extracts were incubated at 37℃for 60h. Finally, the fermented CMBP was boiled for 5min, the LAB was sterilized and then cooled to room temperature. It was used as such as a sample or air-dried at 60℃for 2 days until a powder product was obtained.

PH and total titratable acidity pH was measured using a pH meter (Istek, head, korea). The 10mL sample was homogenized with 90mL deionized water. The total Titratable Acidity (TA) was determined by titration with 0.1M NaOH to a final pH of 8.3.

The reducing sugar content adopts a DNS method (7) as a standard scheme for determining the reducing sugar content. After 0.5mL of sample was mixed with 1.5mL of DNS reagent, the mixture was heated at 100deg.C for 10min. 8mL of distilled water was added to the sample as the temperature was reduced to room temperature. The absorbance at 540nm was determined using a spectrophotometer (V-530 UV/VIS; jasco, tokyo, japan). Calibration curves were made using glucose solutions of different concentrations (0.4-1.5 mg/mL).

Amino nitrogen (amino-N) content the amino nitrogen content was determined according to Nissen method (8). 5mL of the sample was mixed with 45mL of distilled water, and then centrifuged at 7000 Xg for 10min. 0.125mL of the supernatant was mixed with 2mL of 0.2125M sodium phosphate buffer (pH 8.2) and 1mL of 0.01% trinitrobenzenesulfonic acid (TNBS) for 30min. The reaction was terminated by adding 2mL of 0.1N Na ₂SO₃. The absorbance at 420nm was measured.

Microbiological analysis 25mL of the samples were aseptically transferred to sterile plastic bags and homogenized with 225mL of sterile peptone water in a stomach (stomach) for 1min. Suitable decimal dilution samples were prepared using the same diluents and 0.1mL of each diluent was placed in triplicate on different growth media. The following media were used: (1) LAB counts were performed by incubating MRS and BHI agar for 1-2 days at 37 ℃; (2) PCA agar was incubated at 37℃for 1-2 days for total viable cell count. Results are expressed as colony forming units/mL (CFU/mL).

Gas chromatography-Mass Spectrometry (GC-MS) analysis the fermented CMBP volatile compounds were analyzed using a gas chromatography-Mass Spectrometry (GC-MS) (6890N/MSD 5973; agilent Technologies, SANTA CLARA, CA, USA) equipped with CTC CombiPAL auto sampler system, solid Phase Microextraction (SPME) device (Supelco, bellefonte, pa, USA) and HP-5MS capillary column (30 m 0.25mm 0.25 m) (Agilent Technologies). Each sample bottle was left under stirring (500 rpm) at 50 ℃ for 30min for equilibration, then the septum was pierced using an SPME needle. The fibers were exposed to the sample headspace for 30min. Immediately after extraction, the sample was transferred to the sample inlet of the GC and desorbed for 1min at 230 ℃. The oven temperature program is as follows: 5s at 50 ℃; heating to 270 ℃ at the speed of 4 ℃ per second, and preserving heat for 30s. Volatile compounds were identified by comparing their mass spectra to a reference database (version 98 NIST mass spectra data). Only compounds with a similarity of more than 80% (maximum similarity 100) were selected.

Electronic Nose (E-Nose) analysis uses HERACLES E-Nose (Alpha M.O.S., toulouse, france) to distinguish the odor patterns of the different samples. It consists of a headspace sampling system and a detector system (comprising a bipolar (DB-5) and micropolarized (DB-1701) bipost coupled to two flame ionization detectors (MXT-5-FID 1, MXT-1701-FID 2), which allows for clearer identification of compounds. 10mL of the fermented CMBP extract was placed in a 20mL headspace bottle and then capped with a septum-sealed screw cap. The vials were placed in an autosampler of the headspace system. Each flask was incubated at 50deg.C with stirring (500 rpm) for 30min to accumulate volatile compounds in the sample. The gas accumulated in the headspace is then injected into the electronic nose system. The oven temperature program is as follows: 5s at 50 ℃; heating to 270 ℃ at the speed of 4 ℃ per second, and preserving heat for 30s. The two FIDs were at 280 ℃. At least triplicate each analysis. The spectrum of the volatile compounds detected was studied initially by comparing the peaks obtained in the double column, using the Kovats retention index. All obtained data were analyzed using Alpha MOS proprietary software (Alpha m.o.s.).

Amino acid composition 50g of the sample was extracted and hydrolyzed with 10mL 6N HCl at 110℃for 24h. The hydrolyzed solution was evaporated to the dry point using a vacuum evaporator (Buchi Labortechnik, postfach, switzerland). The dried sample was dissolved in 1mL of distilled water and filtered using a 0.2 μm cellulose acetate needle filter (Toyo Roshi Kaisha ltd., tokyo, japan). The resulting solution was analyzed using an amino acid analyzer (Hitachi, tokyo, japan).

Sensory evaluation to evaluate the sensory properties of CMBP extracts and CMBP-LB complex flavourings, 20 panelists (10 men and women each) aged between 23-27 were recruited. For CMBP extracts, 20mL of each sample was filled in a glass beaker and randomly provided to panelists in a sensory chamber at 21+ -1.2 ℃. Referring to the previous report (9), four sensory attributes were used, namely rancid, sour, fishy, grass. The preference scale for the five-point system is used according to the following criteria: 1 (very weak), 2 (quite weak), 3 (neither weak nor strong), 4 (quite strong) and 5 (very strong). For CMBP-LB complex flavourings, four sensory attributes were evaluated, namely taste, smell, colour and overall appearance. The preference scale for the five-point system is used according to the following criteria: 1 (very poor), 2 (quite poor), 3 (neither poor nor good), 4 (quite good) and 5 (very good).

Statistical analysis all experiments were repeated three times. Data are expressed as mean.+ -. SD and analyzed using SPSS software (version 11.0; SPSS Inc., chicago, IL, USA) according to Ducan multiple comparison test (p.ltoreq.0.05).

The experimental results are as follows:

The CMBP powder was analyzed for crude protein, crude fat, ash and carbohydrate content of 23.7% ± 2.2%, 1.4 ± 0.7%, 11.0 ± 1.5% and 63.9 ± 7.2%, respectively. As can be seen, CMBP is rich in carbohydrates and proteins and is therefore presumably useful for microbial fermentation.

The total viable cell count of the fermentation CMBP and control increased to 12h of fermentation, after which it remained almost unchanged (fig. 1). The number of LAB in the fermentation CMBP was slowly decreased after 12h,24h, while the control did not grow LAB during the fermentation. The number of CMBP LABs for 24h of fermentation reached 1.1X10 ⁶ CFU/mL, whereas the number of control LABs was undetectable.

The pH, TA, reducing sugar content of the fermentations CMBP at different fermentation periods varied as shown in FIG. 2. As the fermentation proceeds, the pH of the fermentation CMBP drops more significantly than the control, while TA increases. At 60h of fermentation, the pH of the fermented CMBP was reduced from 6.79 to 4.96-4.87, while TA increased from 0.13 to 0.80-0.71.LAB fermentation CMBP has good effects on pH decrease and TA increase. The reducing sugar content is continuously reduced during the fermentation process. It is therefore believed that microorganisms may use CMBP carbohydrate as a carbon source (10).

The amino nitrogen content of the fermented CMBP is shown in figure 3. The amino nitrogen content of fermented seafood is regarded as a flavor and taste index compound and thus is used to evaluate the quality of fermented seafood (11). After 24h of fermentation, the amino nitrogen content of the control decreased, whereas CMBP fermented by LAB began to decrease after 48h of fermentation. Riebroy et al (12) report that an increase in amino nitrogen may be associated with an increase in the formation or taste of flavor compounds. Thus, fermentation using LAB helps to improve the flavor of the fermented CMBP. The highest point of amino nitrogen content is generally considered to be the optimal fermentation period (13) for the flavor of the fermented food product. Since the amino nitrogen content of the fermented CMBP was highest at 48h fermentation, 48h fermentation was considered to be the optimal fermentation period and CMBP of 48h fermentation was used for further investigation.

Table 1 shows 17 volatile compounds identified and categorized as aldehydes, alkanes, acids, and ketones. Wherein the proportion of ketone is the largest and comprises 40% of the total volatile compounds in the control. Propanal and dodecanal (having a pungent odor and being a component of citrus oil) were also detected in the control (14). By fermentation, the propionaldehyde content in the LB sample is significantly reduced, and the propionaldehyde content in the other fermented CMBP extracts disappears. Furthermore, in all the fermented CMBP extracts, dodecanal completely disappeared. Beta-ionone and 3-buten-2-one are known to cause violet-like odors and seaweed odors (15, 16). By fermentation, beta-ionone and 3-buten-2-one are completely disappeared, while other ketones such as 2-undecanone, 2-nonone, 6-tridecanone and 2-tetradecanone are newly formed in the fermented CMBP extract. Carbonyl-retaining compounds (e.g., aldehydes and ketones) are formed from the degradation of amino acids. Partial degradation of amino acids is associated with the formation of higher alcohols, which can then be converted to aldehydes and ketones in the presence of oxygen (17). Among these ketone compounds, 2-undecanone is known to have a mild and intense sweet fragrance and taste (17). Heptanoic acid is an oily liquid (18) having an unpleasant rancid odor. Dibutyl phthalate has a paint-like smell (16). However, these materials fall to undetectable levels through the fermentation process. In addition, the content of acrylic acid associated with cheese flavor was slightly increased in both the LL and LP samples. Heptadecane (derived from fatty acids, known to have off-flavors (14) in water) in the fermented CMBP extract was significantly reduced to undetectable levels. Thus, by fermentation, the content of off-flavor compounds is reduced or eliminated, similar to the result of Song et al (5). Thus, the use of LAB fermentation to remove seaweed off-flavors can potentially be applied to develop more value added products.

Nd: not found

* Analysis of volatile Compounds Using GC-MS

* Peak area

The volatile compound spectra and Principal Component Analysis (PCA) using E-Nose are shown in Table 2 and FIG. 4, respectively. In comparison to GC-MS data, 5 volatile compounds (3-buten-2-one, E-3-octen-2-one, 3, 5-octen-2-one, 1-butylamine and 1-methoxy-2-propanol) were identified using E-Nose analysis. The differences in GC-MS data and E-Nose data may be caused by different systems and authentication methods. E-Nose consists of a dual column coupled to a dual detector, using retention index to identify unknown compounds, while GC-MS is identified based on nonpolar columns and mass spectrometry. Furthermore, the panelists detected fishy smell according to sensory evaluation (table 4). This is probably due to the presence of 1-butylamine (detected only in E-Nose but not in GC-MS). Thus, it can be concluded that GC-MS in combination with E-Nose can improve the accuracy of identification of volatile compounds. The 99.92% data variability on the horizontal axis and the 0.05445% data variability on the vertical axis, based on the amount of off-flavor compounds (mainly from 1-butylamine), account for the differences in fragrance pattern between samples that are distinguishable on the PC1 axis (fig. 4). In the figure, the closer to the left, the higher the content of the odorous compound, and the closer to the right, the lower the content of the odorous compound. Based on this result, the fermented CMBP extract and control can be distinguished by differences in major off-flavors using the E-Nose system in combination with the Principal Component Analysis (PCA) method. It follows that the malodour of CMBP extracts can be significantly eliminated by LAB fermentation.

The amino acid composition of the food material is important for the nutritional value and has an influence on the functional properties (19). Aspartic acid, glutamic acid and cysteine impart salty taste; methionine, isoleucine and leucine impart a bitter taste; threonine, proline, serine, glycine, alanine and lysine impart sweetness (20). The amino acid content of CMBP extract is shown in table 3. The control is rich in glutamic acid, aspartic acid and glycine, accounting for 15.5%, 12.2 and 11.4% of the amino acid composition, respectively. After fermentation, the content of 7 amino acids (glutamic acid, aspartic acid, threonine, serine, cysteine, methionine and arginine) was reduced in all the fermented CMBP extracts, and the content of amino acids was increased except tyrosine (reduced in LB samples after fermentation).

The control sample consisted of 39.9% sweet amino acids, 28.6% salty amino acids, and 10.6% bitter amino acids. After fermentation, sweet amino acids increased significantly (LB, LL, LP and ST were 46.2, 45.0, 44.9 and 44.6, respectively), while salty amino acids decreased (LB, LL, LP and ST were 24.4, 22.4, 23.6 and 22.3, respectively) and bitter amino acids increased (LB, LL, LP and ST were 11.8, 11.2, 10.9 and 11.7, respectively). Since sweet amino acids increase after fermentation and off-flavor compounds decrease or disappear (tables 2 and 3), it is believed that the fermented CMBP extract can be developed as a raw material for food supplements.

Sensory evaluation of CMBP extracts is shown in table 4. The control samples had a flavor profile of 1.75.+ -. 0.44 (sour) with a malodor of 3.30.+ -. 0.47 (rancid), 3.00.+ -. 0.65 (fishy), 4.40.+ -. 0.50 (grassy), respectively. After fermentation, the fermented CMBP extract had reduced off-flavors (LB, LL, LP and ST, rancid of 2.55.+ -. 0.44, 2.15.+ -. 0.81, 2.55.+ -. 0.60, 2.65.+ -. 0.49, fishy smell of 1.95.+ -. 0.39, 2.00.+ -. 0.32, 1.85.+ -. 0.37, 1.90.+ -. 0.31, grass smell of 2.25.+ -. 0.64, 3.30.+ -. 0.57, 3.10.+ -. 0.64, 3.10.+ -. 0.55), respectively), while the flavor profile (LB, LL, LP and ST, sour smell of 3.50.+ -. 0.51, 3.00.+ -. 0.56, 3.55.+ -. 0.51, 3.20.+ -. 0.52) was increased, respectively. Therefore, LAB fermentation is effective in removing off-flavors, especially in CMBP extracts fermented with LB, which have the highest sensory scores. The reduction in off-taste may be due to the reduction in the corresponding compounds identified by GC-MS and E-Nose analysis, based on the sensory scores of the fermented CMBP extracts, which may be effective in removing off-taste by microbial fermentation (21).

Example 2

This example compares the sensory evaluation differences between a flavor comprising Lactobacillus bulgaricus fermented CMBP (CMBP-LB) and an existing commercial product.

Using CMBP-LB as a raw material, a natural composite seasoning was prepared by mixing: 15g CMBP-LB, 10g solar salt, 2.5g nucleic acid, 5g glucose, 2.5g red pepper powder, 0.5g onion powder, 0.25g ginger powder, 1g garlic powder, 1.5g bean paste powder. For sensory evaluation, 30g CMBP-LB complex flavor was added to 70mL of water, followed by boiling for 5min. After this time, it was cooled to room temperature and used as a sample for sensory analysis. Three evaluations were performed under the same analytical conditions using commercial anchovy compound sauce and beef compound sauce as control. In Table 5, the CMBP-LB complex flavor was not significantly different from the other commercial products in terms of each sensory attribute (p < 0.05). Therefore, CMBP-LB is considered to be useful as an additive in the food industry for more value-added raw materials.

Table 5: sensory evaluation of CMBP-LB Complex flavoring

^a The mean values with different superscripts in the same column are significantly different (p < 0.05).

1) CMBP-LB, a bracket algae byproduct fermented by lactobacillus bulgaricus, a compound seasoning 2) ACS, an anchovy compound seasoning

3) BCS (binary coded decimal) and beef compound seasoning

Thus, CMBP extracts using LAB fermentation may be an alternative method to increase the availability of seaweed by-products for developing biologically functional food materials. The TA and amino nitrogen content in the fermentation CMBP increased significantly, while the pH and reducing sugar content decreased. LAB fermentation significantly removes or reduces off-flavor compounds. The use of PCA allows complete discrimination of fragrance patterns based on the content of odoriferous compounds. The fermented CMBP extract has reduced salty amino acid content, and after fermentation, increased sweet and bitter amino acid content. Based on sensory evaluation, fermented CMBP (especially fermented by lactobacillus bulgaricus) can be developed as a food supplement raw material for natural complex flavors.

The use of lactobacillus fermentation in the preparation of a seasoning is described in detail above, and specific examples are applied herein to illustrate the principles and embodiments of the invention, and the above examples are only for helping to understand the method and core idea of the invention; meanwhile, as those skilled in the art will have variations in the specific embodiments and application scope in accordance with the ideas of the present invention, the present description should not be construed as limiting the present invention in view of the above.

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Claims

1. Use of a lactobacillus-fermented tannery algae processing byproduct in the preparation of a seasoning, characterized in that the lactobacillus is lactobacillus bulgaricus, lactobacillus plantarum, lactococcus lactis or streptococcus thermophilus; fermenting the tannery algae processing byproduct with the lactic acid bacteria for 48 hours; the flavoring agent also comprises solar salt, nucleic acid, glucose, red pepper powder, onion powder, ginger powder, garlic powder and bean paste powder; in the seasoning, the mass ratio of the lactobacillus-fermented tannery algae processing byproducts, solar salt, nucleic acid, glucose, red pepper powder, onion powder, ginger powder, garlic powder and bean paste powder is 60:40:10:20:10:2:1:4:6.

2. The use according to claim 1, characterized in that the lactic acid bacteria are lactobacillus bulgaricus.

3. A seasoning, characterized in that the seasoning comprises a stannum algae processing byproduct fermented by lactic acid bacteria, wherein the lactic acid bacteria are lactobacillus bulgaricus, lactobacillus plantarum, lactococcus lactis or streptococcus thermophilus; fermenting the tannery algae processing byproduct with the lactic acid bacteria for 48 hours; the flavoring agent also comprises solar salt, nucleic acid, glucose, red pepper powder, onion powder, ginger powder, garlic powder and bean paste powder; in the seasoning, the mass ratio of the lactobacillus-fermented tannery algae processing byproducts, solar salt, nucleic acid, glucose, red pepper powder, onion powder, ginger powder, garlic powder and bean paste powder is 60:40:10:20:10:2:1:4:6.

4. A seasoning according to claim 3 wherein the lactic acid bacteria is lactobacillus bulgaricus.