CN109694828A

CN109694828A - The comprehensive processing technique of threonine mother liquor

Info

Publication number: CN109694828A
Application number: CN201811581412.3A
Authority: CN
Inventors: 唐永强; 李德衡; 马延和; 王健; 刘元涛; 刘海涛; 董力青; 李航
Original assignee: Qiqihar Longjiang Fufeng Biotechnology Co ltd; Jilin University; Tianjin Institute of Industrial Biotechnology of CAS
Current assignee: Qiqihar Longjiang Fufeng Biotechnology Co ltd; Jilin University; Tianjin Institute of Industrial Biotechnology of CAS
Priority date: 2018-12-24
Filing date: 2018-12-24
Publication date: 2019-04-30

Abstract

The invention belongs to the field of biotechnology, and discloses a comprehensive treatment process for threonine mother liquor, which comprises the following steps: step 1) preparing fermentation broth of Candida utilis and Lactobacillus casei, step 2) preparing algal bloom Algal liquid, step 3) mother liquor treatment. The process of the invention does not require additional medium components, the process is simple, the cost is low, and it is suitable for processing large-scale mother liquor.

Description

The comprehensive processing technique of threonine mother liquor

Technical field

The invention belongs to field of biotechnology, and in particular to the comprehensive processing technique of threonine mother liquor.

Background technique

China is amino acids production and consumption big country, and often discharge largely contains in the industry productions such as chemical industry, pharmacy, food The waste liquor of amino acid shows as high COD, high NH containing there are many amino acid, residual sugar, fermentation intermediate product, ammonium salt etc.₃- N's Feature, and in acidity, it after being discharged into water body, would seriously pollute the environment, difficulty of governance is larger.To solve waste water pollution problem, together When waste water resource is efficiently used, in traditional production technology, first mycoprotein in amino acid waste liquor is extracted It is sold to be made into feed, the waste liquor after extracting albumen is concentrated to prepare fertilizer regurgitation feeding agriculture using spray granulation.

Amino acid fermentation waste liquid slurry-spraying pelletizing technique is that high-concentration waste water is effectively treated in amino acids production enterprise, realizes circulation Economy increases the effective way of business economic income, but during fermented waste fluid slurry-spraying pelletizing, produces containing large quantity of moisture With the pollution flue gas of organic matter, environment " secondary pollution " is easily caused.It is normal that there is the flue gas serious peculiar smell to influence the masses around Life, become local government and urgently solve the problems, such as, be long-standing problem fermentation industry development significant technology issues.

Some researches show that can use amino acids industry waste water and produce feeding mycoprotein, such as Chinese patent technology " benefit With the method for the inferior Dbaly yeast of the high feeding Chinese of saliferous amino acid wastewater fermenting and producing " to disclose a kind of recycling amino acid useless The method that water carries out fermenting and producing mycoprotein, this method are prepared for culture medium using waste water, and nutrient media components are high saliferous ammonia Base acid sewage: 125g/L-330g/L, carbon source: 30-100g/L, potassium dihydrogen phosphate (KH₂PO₄): 5-15g/L, yeast powder: 0-10g/ L, magnesium sulfate (MgSO₄): 3-9g/L, calcium carbonate (CaCO₃): 0.75-2g/L, ferrous sulfate (FeSO₄): 1.5-4g/L, culture medium PH is 3.5-7.5, and waste water has been effectively treated in this method, and is prepared for mycoprotein；But there is also some defects for this method: The defects of culture medium raw material higher cost, limited waste water usage amount.

Since waste liquor contains multiple nutritional components, during storage pool memory is put, a large amount of microbial floras can be bred, With the accumulation of storage pool runing time, the dominant microflora for being adapted to the low pH of waste liquor, high salt adverse circumstances is formd in pond. Research and development centre, Fu Feng group had once carried out case study (Li Haiyan, the equal " dominant bacteria in gourmet powder waste water of Xu Guohua to this project The research of separation screening and growth characteristics ", science and technology of fermenting communication, the 4th phase in 2008), the results showed that it is raw in gourmet powder waste water A large amount of microorganism not of the same race is had, under certain ecological condition, the growth of various microorganisms is in equilibrium state；Wherein Dominant growth bacterium remains dominant growth in waste water, and the ratio of microbial count is maximum in water body shared by increment, by grinding Study carefully the external condition for changing dominant bacteria, improves its growth activity, will be greatly improved to the degradation effect of Wastewater Pollutant. As it can be seen that amino acid waste liquid is suitble to the breeding of microorganism, it can be with pollutant in decomposition nut liquid；We can be specific by adding The microorganism of mother liquor environment is adapted to, to decompose the pollutant in span amino acid mother liquor, to achieve the purpose that integrated treatment mother liquor.

Patented technology " CN107267427A, a kind of threonine mother liquor recycling method " before applicant utilizes withered grass Bacillus, bacillus licheniformis and scenedesmus obliquus handle mother liquor, and effect is preferable, but need to withered grass gemma Bacillus, bacillus licheniformis and scenedesmus obliquus are tamed, and need additional addition nutrient media components, complex process, at This raising is unfavorable for handling extensive mother liquor.

Summary of the invention

In order to overcome the existing technological deficiency of the prior art further to be improved, the present invention provides threonine mothers The comprehensive processing technique of liquid.

For achieving the above object, the invention adopts the following technical scheme:

The comprehensive processing technique of threonine mother liquor comprising following steps: step 1) prepares candida utili bacterium and cheese cream The zymocyte liquid of bacillus, step 2 prepare Anabaena Flos-aquae liquid, step 3) mother liquid disposal.

Further,

Step 1) prepares the zymocyte liquid of candida utili bacterium and Lactobacillus casei: taking appropriate threonine mother liquor, 100 DEG C are boiled Then 5min adds ammonium chloride, stir evenly, then is successively inoculated with candida utili bacterium seed liquor and Lactobacillus casei seed Liquid, temperature are 28-32 DEG C, with 200rpm shake culture 12h, are then allowed to stand culture for 24 hours, obtain candida utili bacterium and cheese The zymocyte liquid of lactobacillus；

Step 2 prepares Anabaena Flos-aquae liquid: the Anabaena Flos-aquae in logarithmic growth phase is inoculated into according to 10% inoculum concentration It is cultivated in BG11 culture medium, the condition of culture are as follows: intensity of illumination 5000lux, 25 DEG C of cultures, Light To Dark Ratio 12:12, ventilation Rate is 0.2vvm；48h is cultivated, algae solution is collected；

Step 3) mother liquid disposal: threonine mother liquor is discharged in reaction tank, is then laid with non-woven fabrics as attachment carrier, then will hair Yeast-like fungi liquid is inoculated into reaction tank, and the processing time is 2-3d, and then algae solution is inoculated into reaction tank, continues with 6-7d, plate Microorganism is collected by filtration in frame, is then discharged out liquid, is used for field irrigation.

Preferably,

In the step 1), the additive amount of ammonium chloride accounts for the 0.5-1% parts by weight of threonine mother liquor.

Preferably,

In the step 1), the inoculum concentration of the candida utili bacterium seed liquor and Lactobacillus casei seed liquor is 5-10%.

Preferably,

In the step 3), the inoculum concentration of the zymocyte liquid is 1-2%.

Preferably,

In the step 3), the inoculum concentration of the algae solution is 0.5-1%.

Preferably,

In the step 1), the candida utili bacterium seed liquor the preparation method comprises the following steps: candida utili bacterium is inoculated into It is cultivated in YPD fluid nutrient medium, obtains candida utili bacterium seed liquor.

Preferably,

In the step 1), the Lactobacillus casei seed liquor is trained the preparation method comprises the following steps: Lactobacillus casei is inoculated into MRS liquid It supports and is cultivated in base, obtain Lactobacillus casei seed liquor.

Compared with prior art, the beneficial effect that the present invention obtains mainly includes but is not limited to the following aspects:

Since oxygen content is lower in mother liquor, the present invention is inoculated with candida utili bacterium and Lactobacillus casei first, and two kinds of bacterial strains can With mutual symbiosis, under anaerobic, macromolecular organic matter is decomposed, generate small organic molecule, inhibit miscellaneous bacteria, the later period passes through again Anabaena Flos-aquae is degraded the organic nutrient substances such as nitrogen, phosphorus in mother liquor in the way of small molecule organic matter, also using too Sunlight carries out photosynthesis to guarantee the growth and breeding of itself, absorbing carbon dioxide, and generates oxygen, moreover it is possible to promote bacterial strain raw It is long, entire method simple possible, low in cost, high treating effect；Non-woven fabrics large specific surface area, it is not easy to it is corroded, microorganism It can adhere to form biomembrane, biofilm effect is good, and treatment effeciency is high.

Figure of description

Fig. 1: influence of each factor to COD in mother liquor and ammonia nitrogen removal frank；

Fig. 2: influence of the inoculum concentration of Anabaena Flos-aquae to major pollutants COD and ammonia nitrogen removal frank.

Specific embodiment

In order to make those skilled in the art better understand the technical solutions in the application, having below in conjunction with the application Body embodiment more clearly and completely describes the present invention, it is clear that described embodiment is only the application one Divide embodiment, instead of all the embodiments.Based on the embodiment in the application, those of ordinary skill in the art are not making Every other embodiment obtained, should fall within the scope of the present invention under the premise of creative work.

Embodiment 1

The comprehensive processing technique of threonine mother liquor comprising following steps:

Candida utili bacterium is inoculated into YPD fluid nutrient medium and is cultivated, candida utili bacterium seed liquor, bacterium are obtained Bulk concentration is 1 × 10⁹CFU/ml；

Lactobacillus casei is inoculated into MRS fluid nutrient medium and is cultivated, Lactobacillus casei seed liquor, cell concentration control are obtained System is 3 × 10⁹CFU/ml；

Appropriate threonine mother liquor is taken, 100 DEG C are boiled 5min, and then addition accounts for the ammonium chloride of 0.6% parts by weight of threonine mother liquor, are stirred It mixes uniformly, is then successively inoculated with candida utili bacterium seed liquor and Lactobacillus casei seed liquor, inoculum concentration are 10%, temperature It is 30 DEG C, with 200rpm shake culture 12h, is then allowed to stand culture for 24 hours, obtains zymocyte liquid；

It will be in the Anabaena Flos-aquae of logarithmic growth phase, is inoculated into BG11 culture medium and is cultivated according to 10% inoculum concentration, Inoculum density is 1 × 10⁵A/ml, the condition of culture are as follows: intensity of illumination 5000lux, 25 DEG C of cultures, Light To Dark Ratio 12:12 lead to Gas velocity rate is 0.2vvm；48h is cultivated, algae solution is collected；

Threonine mother liquor is discharged in reaction tank, non-woven fabrics is then laid with as attachment carrier, will be sent out according still further to 2% inoculum concentration Yeast-like fungi liquid is linked into reaction tank, and the processing time is 2d, and then algae solution is linked into reaction tank according to 1% inoculum concentration, is continued 6d is handled, plate-frame filtering collects microorganism, is then discharged out liquid, is used for field irrigation.

Embodiment 2

Candida utili bacterium is inoculated into YPD fluid nutrient medium and is cultivated, candida utili bacterium seed liquor, bacterium are obtained Bulk concentration is 2 × 10⁹CFU/ml；

Lactobacillus casei is inoculated into MRS fluid nutrient medium and is cultivated, Lactobacillus casei seed liquor, cell concentration control are obtained System is 5 × 10⁹CFU/ml；

Appropriate threonine mother liquor is taken, 100 DEG C are boiled 5min, and then addition accounts for the ammonium chloride of 0.5% parts by weight of threonine mother liquor, are stirred It mixes uniformly, is then successively inoculated with candida utili bacterium seed liquor and Lactobacillus casei seed liquor, inoculum concentration are 6%, temperature is It 28 DEG C, with 200rpm shake culture 12h, is then allowed to stand culture for 24 hours, obtains zymocyte liquid；

Threonine mother liquor is discharged in reaction tank, non-woven fabrics is then laid with as attachment carrier, will be sent out according still further to 1% inoculum concentration Yeast-like fungi liquid is linked into reaction tank, and the processing time is 3d, and then algae solution is linked into reaction tank according to 0.5% inoculum concentration, after Continuous processing 7d, plate-frame filtering collect microorganism, are then discharged out liquid, are used for field irrigation.

Embodiment 3

The threonine mother liquor that the present invention uses is the raffinate after extracting threonine in fermentation liquid, randomly selects 2018 Nian Sanji Certain a batch of threonine mother liquor of degree is tested, before detecting threonine mother liquid disposal, treated each pollutant index, tool Body is shown in Table 1:

Table 1

Index	Before processing	After processing (1 technique of embodiment)
			COD(mg/L)	3218.5	53.1
Ammonia nitrogen (mg/L)	416.2	12.8
			SS(mg/L)	157.3	7.9
P(mg/L)	113.8	5.7
			Crude protein (g/L)	29.6	4.1

As shown in Table 1, after treatment, the pollutant in mother liquor and the crude protein content range of decrease are obvious, and water quality completely can be with For field irrigation water source, energy-saving and emission-reduction reduce the environmental protection pressure of enterprise.

Embodiment 4

Using major pollutants COD and ammonia nitrogen as Testing index, influence of each factor to mother liquid disposal is verified.

Different control groups is set, wherein control 1: does not add candida utili bacterium, remaining is the same as embodiment 1；Control 2: Lactobacillus casei is not added, remaining is the same as embodiment 1；Control 3: Anabaena Flos-aquae is not added, remaining is the same as embodiment 1；Control 4: three Kind microorganism is accessed simultaneously, remaining is the same as embodiment 1.Experimental group is embodiment.As shown in Figure 1, compared with control 1-4, experiment Group is broken down into small molecule to part larger molecular organics, then using candida utili bacterium and Lactobacillus casei is first accessed The mode of Anabaena Flos-aquae is accessed, Anabaena Flos-aquae proliferative speed is improved, can be improved the removal rate of major pollutants in mother liquor, Effect is better than control 1-4.

Embodiment 5

Influence of the inoculum concentration of Anabaena Flos-aquae to major pollutants COD and ammonia nitrogen removal frank.

The inoculum concentration that inoculum concentration is respectively 0,0.25,0.5,1.0,2.0,4.0 (%) six different gradients is arranged to be examined It surveys, as shown in Fig. 2, COD and ammonia nitrogen removal frank improve rapidly with the increase of inoculum concentration, when increasing to 1%, reaches peak value, Then as the increase of inoculum concentration, COD and ammonia nitrogen removal frank decline, it may be possible to because of Anabaena Flos-aquae excessively fast passage, cause algae Class Mortality and water pollution are unfavorable for the removal of pollutant instead.

It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with A variety of variations, modification, replacement can be carried out to these embodiments without departing from the principles and spirit of the present invention by understanding And modification, the scope of the present invention is defined by the appended.

Claims

1. The comprehensive treatment process of threonine mother liquor, which comprises the following steps: step 1) preparing fermented bacteria liquid of Candida utilis and Lactobacillus casei, step 2) preparing liquid of Anabaena bloom, step 3) mother liquor deal with.

2. comprehensive treatment process according to claim 1, is characterized in that,

Step 1) Prepare the fermentation broth of Candida utilis and Lactobacillus casei: take an appropriate amount of threonine mother solution, boil at 100°C for 5 minutes, then add ammonium chloride, stir evenly, and then inoculate Candida utilis seeds in sequence liquid and Lactobacillus casei seed liquid, the temperature is 28-32 ℃, shake and cultivate at 200rpm for 12h, and then stand for 24h, to obtain the fermentation bacteria liquid of Candida utilis and Lactobacillus casei;

Step 2) Preparation of Anabaena alba solution: Inoculate the Anabaena alba in the logarithmic growth phase into BG11 medium according to 10% of the inoculum for cultivation. The cultivation conditions are: light intensity of 5000 lux, cultivation at 25°C , the light-dark ratio is 12:12, and the ventilation rate is 0.2vvm; after culturing for 48h, the algal fluid is collected;

Step 3) Mother liquor treatment: Discharge the threonine mother liquor into the reaction tank, then lay the non-woven fabric as the attachment carrier, and then inoculate the fermentation bacteria liquid into the reaction tank for 2-3 days, and then inoculate the algae liquid into the reaction tank. In the reaction tank, the treatment is continued for 6-7 days, the microorganisms are collected by plate and frame filtration, and the liquid is finally discharged for field irrigation.

3. The comprehensive treatment process according to claim 2, wherein in the step 1), the addition of ammonium chloride accounts for 0.5-1% by weight of the threonine mother liquor.

4 . The comprehensive treatment process according to claim 2 , wherein in the step 1), the inoculum amounts of the Candida utilis seed solution and the Lactobacillus casei seed solution are both 5-10%. 5 .

5 . The comprehensive treatment process according to claim 2 , wherein, in the step 3), the inoculum of the fermentation bacterial liquid is 1-2%. 6 .

6 . The comprehensive treatment process according to claim 2 , wherein in the step 3), the inoculum of the algal liquid is 0.5-1%. 7 .

7 . The comprehensive treatment process according to claim 2 , wherein in the step 1), the preparation method of the Candida utilis seed liquid is: inoculating the Candida utilis into the YPD liquid. 8 . Cultivated in the medium to obtain Candida utilis seed liquid.

8. The comprehensive treatment process according to claim 2, wherein in the step 1), the preparation method of the Lactobacillus casei seed solution is: inoculating the Lactobacillus casei into the MRS liquid medium for cultivation, Lactobacillus casei seed solution was obtained.