GA3 production extraction wastewater treatment method

Abstract

The invention discloses a GA3 production extraction wastewater treatment method which comprises the following steps: S1, filling GA3 production extraction wastewater into a fermentation container, sterilizing, and inoculating a saccharomyces cerevisiae seed solution with the inoculation amount of 1-15v / v%; s2, performing fermentation culture to obtain fermentation liquor under the fermentation conditions that the temperature is 25-35 DEG C, the speed is 50-600r / min and the time is 24-200 hours. According to the method, valuable single-cell protein is produced, and meanwhile, COD is reduced, so that the purpose of treating wastewater is achieved.

Description

Technical Field

[0001] The invention relates to the field of wastewater treatment, and in particular to a method for treating GA3 production extraction wastewater. Background Art

[0002] A large amount of extraction wastewater is generated during the production of GA3 (gibberellin), which contains a large amount of nutrients such as polypeptides, amino acids, esters, alcohols, trace elements, etc., and the chemical oxygen demand (COD) is as high as 50,000-100,000 mg / L.

[0003] The current treatment of the extracted wastewater generated in the production of GA3 is to directly send the extracted wastewater into the sewage treatment system to undergo chemical, physical and biochemical reactions (biochemical reactions of ordinary wastewater) like ordinary wastewater. On the one hand, this seriously increases the difficulty of subsequent sewage treatment. On the other hand, the extracted wastewater from GA3 production is extremely easy to deteriorate and become smelly, causing serious environmental pollution.

[0004] Therefore, how to effectively treat GA3 production extraction wastewater before sending it into the sewage treatment system has become an urgent problem that the industry needs to solve.

[0005] The above background technology is for facilitating the understanding of the present invention and is not the known technology disclosed to the general public before the application of the present invention. Summary of the invention

[0006] In view of the above-mentioned defects, the present invention provides a method for treating GA3 production extraction wastewater, which produces valuable single-cell protein while reducing its COD to achieve the purpose of treating wastewater.

[0007] The technical solution is: a method for treating GA3 production extraction wastewater, comprising the following steps: S1, put GA3 production extraction wastewater into the fermentation container, sterilize it, and inoculate it with brewer's yeast seed liquid, with an inoculation amount of 1-15v / v; S2, fermentation culture to obtain fermentation liquid, the fermentation conditions are: 25~35℃, 50~600r / min, 24~200h.

[0008] Furthermore, the brewer's yeast is Angel highly active dry yeast.

[0009] Furthermore, in S2, the fermentation conditions were: 30~33°C, 200~300r / min, 96~120h, pH 3.2~5.5; in S1, the inoculation amount was 5~10 v / v%.

[0010] Furthermore, 0.5-2 g / L potassium dihydrogen phosphate is added to S1.

[0011] Furthermore, in S1, the fermentation container is a shake flask.

[0012] Furthermore, S2 further includes S3: concentrating and drying the fermentation liquid of S2 to obtain a single cell protein product Furthermore, in S1, the fermentation container is a fermentation tank with a charge factor of 50-80%, and in S2, the actual digestion time is 20-50 min and the ventilation volume is 0.5-2 vvm.

[0013] Furthermore, after S2, S3 is also included: separating the fermentation liquid of S2 into a concentrated liquid and a diluted liquid through a membrane concentration, spray-drying the concentrated liquid to obtain a single-cell protein product, and the diluted liquid is wastewater, and the wastewater is discharged into a sewage treatment system.

[0014] Furthermore, the brewer's yeast seed solution is formed by the following steps: S01, YPD plate culture: After the YPD medium was sterilized, the Saccharomyces cerevisiae strain was streaked on a YPD plate for activation and cultured at 25-35°C for 12-48 h; S02, shake flask culture: pick a single colony from the YPD plate and inoculate it into a sterile YPD liquid shake flask, shake it at 25~35℃, 100~300r / min for 12~48h.

[0015] Furthermore, the brewer's yeast seed solution is formed by the following steps: S01, YPD plate culture: After the YPD medium was sterilized, the Saccharomyces cerevisiae strain was activated by streaking on a YPD plate and cultured at 25-35°C for 12-48 h; S02, shake flask culture: pick a single colony from the YPD plate and inoculate it into a sterilized YPD liquid shake flask, shake it at 25-35°C, 100-300 r / min and culture it for 12-48h; S03, seed tank culture: add seed tank culture medium into 15L seed tank, filling factor 50~80%, actual sterilization 20~50min, inoculation amount 1~5v / v%, ventilation volume 100~300r / min, ventilation volume 0.5~1vvm, culture at 25~35℃ for 12~48h.

[0016] Compared with the prior art, the present invention has the following beneficial effects: The present invention uses GA3 production extraction wastewater as raw material to produce single cell protein, and single cell protein (SCP), also known as microbial protein, bacterial protein, has the characteristics of high protein content, balanced amino acid composition, high nutritional value and safety. It is extremely rich in nutrients, with a protein content of up to 40-80%, and contains a variety of vitamins, lipids, amino acids, trace elements, enzymes and bioactive substances, and can be used as food, feed or food additives. Not only can single cell protein be produced using organic matter in wastewater, but the content of organic matter in sewage can also be reduced, thereby realizing the treatment and purification of extraction wastewater. DETAILED DESCRIPTION

[0017] As used herein: "Prepared from" is synonymous with "comprising." As used herein, the terms "comprising," "including," "having," "containing," or any other variation thereof, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises the listed elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.

[0018] The conjunction "consisting of excludes any unspecified element, step, or component. If used in a claim, this phrase renders the claim closed-ended so that it does not include materials other than those described, except for conventional impurities associated therewith. When the phrase "consisting of" appears in a clause of the body of a claim rather than immediately following the subject matter, it limits only the elements described in that clause; other elements are not excluded from the claim as a whole.

[0019] When an amount, concentration, or other value or parameter is expressed as a range, a preferred range, or a range defined by a series of upper preferred values ​​and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pairing of any range upper limit or preferred value with any range lower limit or preferred value, regardless of whether the range is disclosed separately. For example, when a range of "1 to 5" is disclosed, the described range should be interpreted as including ranges "1 to 4", "1 to 3", "1 to 2", "1 to 2 and 4 to 5", "1 to 3 and 5", etc. When a numerical range is described in this article, unless otherwise stated, the range is intended to include its end values ​​and all integers and fractions within the range.

[0020] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0021] Those skilled in the art will appreciate that the following examples are only used to illustrate the present application and should not be considered to limit the scope of the present application. If specific conditions are not specified in the examples, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents, instruments or microorganisms used is not specified, they are all conventional products that can be purchased commercially.

[0022] In these examples, parts and percentages are by mass unless otherwise indicated.

[0023] "Parts by mass" refers to the basic unit of measurement for expressing the mass ratio of multiple components. 1 part can represent any unit mass, such as 1g or 2.689g. If we say that the mass of component A is a parts and the mass of component B is b parts, it means the ratio of the mass of component A to the mass of component B is a:b. Alternatively, it means that the mass of component A is aK and the mass of component B is bK (K is an arbitrary number, indicating a multiple factor). It should not be misunderstood that, unlike the mass parts, the sum of the mass of all components is not limited to 100 parts.

[0024] “And / or” is used to indicate that one or both of the stated situations may occur, for example, A and / or B includes (A and B) and (A or B).

[0025] In the present invention, OD is the optical density value of the fermentation liquid measured at a wavelength of 570 nm using an ultraviolet-visible spectrophotometer.

[0026] In the following embodiments and comparative examples, the components of the shake flask culture medium are: 1% yeast extract powder, 2% peptone, 2% glucose, and the balance is water; the components of the seed tank culture medium are: 1wt% yeast powder, 2wt% peptone, 2wt% glucose, 0.05wt% folate, and the balance is water.

[0027] In the following examples and comparative examples, the initial COD of the extracted wastewater was 73468 mg / L.

[0028] In the following examples, the brewer's yeast was 500 g of Angel High Activity Dry Yeast - Low Sugar Type (Angel Yeast Co., Ltd.).

[0029] Example 1 Preparation of Saccharomyces cerevisiae seed solution, Yarrowia lipolytica seed solution, and Candida albicans seed solution S01, YPD plate culture: After the YPD medium was sterilized, the Saccharomyces cerevisiae, Yarrowia lipolytica, and Candida albicans strains were streaked on YPD plates for activation and cultured at 30°C for 24 h; S02, shake flask culture: single colonies were picked from the YPD plate and inoculated into sterilized YPD liquid shake flasks, and cultured at 30℃ and 200r / min for 24h; The seed liquid of Saccharomyces cerevisiae, the seed liquid of Yarrowia lipolytica and the seed liquid of Candida albicans were obtained respectively.

[0030] Example 2 A method for treating GA3 production extraction wastewater comprises the following steps: S1, put the GA3 production extraction wastewater into a fermentation shake flask, sterilize it, and inoculate the brewer's yeast seed solution of Example 1 at an inoculation amount of 10v / v%.

[0031] S2, culture at 30℃ and 200r / min for 114h, pH natural (3.2-3.5), obtain fermentation broth, and detect OD and COD contents in the fermentation broth. The results are shown in Table 1.

[0032] S3, the fermentation liquid is concentrated and dried to obtain single cell protein powder. The single cell protein content is shown in Table 1, wherein the crude protein content is shown in Table 1.

[0033] Example 3 The difference between this embodiment and embodiment 2 is that in this embodiment, in S2, the culture temperature is 33°C.

[0034] The OD and COD contents in the fermentation broth were detected, and the results are shown in Table 1; The single cell protein yield and crude protein content are shown in Table 1.

[0035] Example 4 The difference between this embodiment and embodiment 2 is that in this embodiment, the pH value of S2 is 5.5.

[0036] The OD and COD contents in the fermentation broth were detected, and the results are shown in Table 1; The single cell protein yield and crude protein content are shown in Table 1.

[0037] Comparative Example 1 The only difference between this comparative example and Example 2 is that in this comparative example, the seed solution of Yarrowia lipolytica in Example 1 is inoculated in S1.

[0038] The OD and COD contents in the fermentation broth were detected, and the results are shown in Table 1; The single cell protein yield and crude protein content are shown in Table 1.

[0039] Comparative Example 2 The only difference between this comparative example and comparative example 1 is that in this comparative example, in S2, the culture temperature is 33°C.

[0040] The OD and COD contents in the fermentation broth were detected, and the results are shown in Table 1; The single cell protein yield and crude protein content are shown in Table 1.

[0041] Comparative Example 3 The only difference between this comparative example and comparative example 1 is that in this comparative example, the pH value of S2 is 5.5.

[0042] The OD and COD contents in the fermentation broth were detected, and the results are shown in Table 1; The single cell protein yield and crude protein content are shown in Table 1.

[0043] Comparative Example 4 The only difference between this comparative example and Example 2 is that in this comparative example, the Candida seed solution of Example 1 is inoculated in S1.

[0044] The OD and COD contents in the fermentation broth were detected, and the results are shown in Table 1; The single cell protein yield and crude protein content are shown in Table 1.

[0045] Comparative Example 5 The only difference between this comparative example and comparative example 4 is that in this comparative example, in S2, the culture temperature is 33°C.

[0046] The OD and COD contents in the fermentation broth were detected, and the results are shown in Table 1; The single cell protein yield and crude protein content are shown in Table 1.

[0047] Comparative Example 6 The only difference between this comparative example and comparative example 4 is that in this comparative example, the pH value of S2 is 5.5.

[0048] The OD and COD contents in the fermentation broth were detected, and the results are shown in Table 1; The single cell protein yield and crude protein content are shown in Table 1.

[0049] Example 5 S01, YPD plate culture: After the YPD medium was sterilized, the Saccharomyces cerevisiae strain was streaked on the YPD plate for activation and cultured at 30°C for 24 h; S02, shake flask culture: single colonies were picked from the YPD plate and inoculated into sterilized YPD liquid shake flasks, and cultured at 30℃ and 200r / min for 24h; S03, seed tank culture: add the seed tank culture medium into a 15L seed tank, with a filling factor of 70%, and sterilize for 30 minutes. Inoculate the shake flask seeds of S02 with an inoculation amount of 2v / v%, aeration of 0.7vvm at 200r / min, and culture at 30℃ for 24h.

[0050] Obtain Saccharomyces cerevisiae seed liquid.

[0051] Example 6 A method for treating GA3 production extraction wastewater comprises the following steps: S1, GA3 production extraction wastewater is charged into a fermentation tank (50L), sterilized, and then inoculated with the brewer's yeast seed solution of Example 5, with a charging factor of 70% and an inoculation amount of 8-10%.

[0052] S2, actual sterilization for 30 min, 300 r / min, ventilation volume 0.7 vvm, defoaming agent automatically controls defoaming, pH is not naturally regulated, culture at 30 ° C for 114 h, and obtain fermentation liquid.

[0053] S3, the fermentation liquid is separated into concentrated liquid and dilute liquid through membrane concentration, and the concentrated liquid is spray-dried to obtain single-cell protein product (the results are shown in Table 1, where the crude protein content is shown in Table 1), and the dilute liquid is wastewater, which is discharged into the sewage treatment system. The COD of the discharged wastewater is 21239 mg / L The OD and COD contents in the fermentation broth were detected. The results are shown in Table 1.

[0054] Example 7 The difference between this embodiment and embodiment 6 is that: in S1, 1g / L potassium dihydrogen phosphate is also added to the fermentation tank (calculated based on a loading factor of 70%, the added amount is 35g). The COD of the discharged wastewater is 18205mg / L The OD and COD contents in the fermentation broth were detected, and the results are shown in Table 1; The single cell protein yield and crude protein content are shown in Table 1.

[0055] Table 1 .

[0056] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for treating GA3 production extraction wastewater, characterized in that: The following steps are involved: S1, put GA3 production extraction wastewater into the fermentation container, sterilize it, and inoculate it with brewer's yeast seed liquid, with an inoculation amount of 1-15v / v; S2, fermentation culture to obtain fermentation liquid, the fermentation conditions are: 25~35℃, 50~600r / min, 24~200h.

2. The GA3 production extraction wastewater treatment method according to claim 1, characterized in that: The brewer's yeast is Angel high-activity dry yeast.

3. The GA3 production extraction wastewater treatment method according to claim 2, characterized in that: In S2, the fermentation conditions were: 30~33℃, 200~300r / min, 96~120 h, pH 3.2~5.5; in S1, the inoculation amount was 5~10 v / v%.

4. The method for treating GA3 production extraction wastewater according to claim 1, characterized in that: In S1, 0.5~2g / L potassium dihydrogen phosphate is also added.

5. The method for treating GA3 production extraction wastewater according to claim 1, characterized in that: In S1, the fermentation container is a shake flask.

6. The method for treating GA3 production extraction wastewater according to claim 5, characterized in that: S2 also includes S3: concentrating and drying the fermentation liquid of S2 to obtain a single cell protein product.

7. The method for treating GA3 production extraction wastewater according to claim 1, characterized in that: In S1, the fermentation container is a fermentation tank with a filling factor of 50-80%. In S2, the actual fermentation time is 20-50 min and the ventilation volume is 0.5-2 vvm.

8. The method for treating GA3 production extraction wastewater according to claim 7, characterized in that: S2 also includes S3: the fermentation liquid of S2 is separated into a concentrated liquid and a dilute liquid through a membrane concentration, the concentrated liquid is spray-dried to obtain a single-cell protein product, and the dilute liquid is waste water, which is discharged into a sewage treatment system.

9. The method for treating GA3 production extraction wastewater according to claim 1, characterized in that: In S1, the saccharomyces cerevisiae seed solution is formed by the following steps: S01, YPD plate culture: After the YPD medium was sterilized, the Saccharomyces cerevisiae strain was streaked on a YPD plate for activation and cultured at 25-35°C for 12-48 h; S02, shake flask culture: pick a single colony from the YPD plate and inoculate it into a sterile YPD liquid shake flask, shake it at 25~35℃, 100~300r / min for 12~48h.

10. The method for treating GA3 production extraction wastewater according to any one of claims 1 to 8, characterized in that: The saccharomyces cerevisiae seed solution is formed by the following steps: S01, YPD plate culture: After the YPD medium was sterilized, the Saccharomyces cerevisiae strain was streaked on a YPD plate for activation and cultured at 25-35°C for 12-48 h; S02, shake flask culture: pick a single colony from the YPD plate and inoculate it into a sterilized YPD liquid shake flask, shake it at 25-35°C, 100-300 r / min and culture it for 12-48 hours; S03, seed tank culture: add seed tank culture medium into 15L seed tank, filling factor 50~80%, actual sterilization 20~50min, inoculation amount 1~5v / v%, ventilation volume 100~300r / min, ventilation volume 0.5~1vvm, culture at 25~35℃ for 12~48h.