CN108330078B - Bacillus licheniformis for improving crude protein yield in guar meal fermentation and application thereof - Google Patents
Bacillus licheniformis for improving crude protein yield in guar meal fermentation and application thereof Download PDFInfo
- Publication number
- CN108330078B CN108330078B CN201710695330.0A CN201710695330A CN108330078B CN 108330078 B CN108330078 B CN 108330078B CN 201710695330 A CN201710695330 A CN 201710695330A CN 108330078 B CN108330078 B CN 108330078B
- Authority
- CN
- China
- Prior art keywords
- fermentation
- guar meal
- bacillus licheniformis
- guar
- crude protein
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 235000012054 meals Nutrition 0.000 title claims abstract description 47
- 238000000855 fermentation Methods 0.000 title claims abstract description 45
- 230000004151 fermentation Effects 0.000 title claims abstract description 45
- 241000194108 Bacillus licheniformis Species 0.000 title claims abstract description 28
- 235000019750 Crude protein Nutrition 0.000 title claims abstract description 18
- 244000007835 Cyamopsis tetragonoloba Species 0.000 title claims abstract 11
- 238000004321 preservation Methods 0.000 claims abstract description 16
- 238000000034 method Methods 0.000 claims abstract description 10
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 4
- 238000009629 microbiological culture Methods 0.000 claims abstract description 4
- 239000001963 growth medium Substances 0.000 claims description 24
- 108090000623 proteins and genes Proteins 0.000 claims description 20
- 239000007788 liquid Substances 0.000 claims description 19
- 102000004169 proteins and genes Human genes 0.000 claims description 19
- 230000001580 bacterial effect Effects 0.000 claims description 16
- 239000002609 medium Substances 0.000 claims description 11
- 238000002156 mixing Methods 0.000 claims description 5
- 235000016709 nutrition Nutrition 0.000 abstract description 6
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 244000303965 Cyamopsis psoralioides Species 0.000 description 40
- 235000018102 proteins Nutrition 0.000 description 18
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 12
- 238000012258 culturing Methods 0.000 description 9
- 244000046052 Phaseolus vulgaris Species 0.000 description 6
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 235000019764 Soybean Meal Nutrition 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 239000004455 soybean meal Substances 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 239000001888 Peptone Substances 0.000 description 3
- 108010080698 Peptones Proteins 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 235000019319 peptone Nutrition 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 229920002907 Guar gum Polymers 0.000 description 2
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 2
- 235000015278 beef Nutrition 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- 239000005018 casein Substances 0.000 description 2
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 2
- 235000021240 caseins Nutrition 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 239000000665 guar gum Substances 0.000 description 2
- 229960002154 guar gum Drugs 0.000 description 2
- 235000010417 guar gum Nutrition 0.000 description 2
- 230000008676 import Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- OMDQUFIYNPYJFM-XKDAHURESA-N (2r,3r,4s,5r,6s)-2-(hydroxymethyl)-6-[[(2r,3s,4r,5s,6r)-4,5,6-trihydroxy-3-[(2s,3s,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]methoxy]oxane-3,4,5-triol Chemical group O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O[C@H]2[C@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)[C@H](O)[C@H](O)[C@H](O)O1 OMDQUFIYNPYJFM-XKDAHURESA-N 0.000 description 1
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 208000031295 Animal disease Diseases 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 241000220485 Fabaceae Species 0.000 description 1
- 229920000926 Galactomannan Polymers 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 108010064851 Plant Proteins Proteins 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 244000098338 Triticum aestivum Species 0.000 description 1
- 101710162629 Trypsin inhibitor Proteins 0.000 description 1
- 229940122618 Trypsin inhibitor Drugs 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003674 animal food additive Substances 0.000 description 1
- 230000000433 anti-nutritional effect Effects 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 210000004913 chyme Anatomy 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000013325 dietary fiber Nutrition 0.000 description 1
- 235000019621 digestibility Nutrition 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000012262 fermentative production Methods 0.000 description 1
- 210000003495 flagella Anatomy 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 235000021118 plant-derived protein Nutrition 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 229940054269 sodium pyruvate Drugs 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 239000002753 trypsin inhibitor Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Polymers & Plastics (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Physiology (AREA)
- Molecular Biology (AREA)
- Animal Husbandry (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Botany (AREA)
- Mycology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses bacillus licheniformis for improving the yield of crude protein in guar meal fermentation, wherein the preservation number of the bacillus licheniformis (Bacillus lincheniformis) is CGMCC No. 14185; the preservation unit is as follows: china general microbiological culture Collection center; the address of the preservation unit is as follows: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North; the preservation date is as follows: 24/05/2017. The invention also provides application of the strain in guar meal fermentation and a fermentation method for improving the crude protein yield of guar meal. The bacillus licheniformis provided by the invention can improve the crude protein yield of guar meal fermentation, and is beneficial to improving the quality and the nutritional value of feed.
Description
Technical Field
The invention relates to the technical field of microbial fermentation, in particular to bacillus licheniformis for improving the crude protein content of guar meal and application thereof.
Background
Guar meal (guar meal) is a by-product produced by degumming guar and then leaching, guar (cyamopsistragonoloba) is a member of the genus guar of the family leguminosae, and is an annual herbaceous drought-resistant crop. Guar meal is rich in protein and carbohydrate, and is a non-transgenic product which is purely natural and free of any chemicals and preservatives. The protein mass fraction of the feed can reach 48-55.16%, and meanwhile, the amino acid contained in the feed is lacked in corn protein, wheat protein and rice protein, and because the guar bean pulp is rich in nutrition and much lower in price than the bean pulp, the feed can better improve the content of the feed protein when being used as a novel high-protein bean pulp raw material. In addition, guar meal also has dietary fiber effect and functional oligosaccharide effect, wherein the maximum crude fiber content is 6.8%, while the crude fiber content in common soybean meal is only 3%.
Guar gum is remained in the guar meal, and the main component of the guar gum is galactomannan, so that the viscosity of chyme in the digestive tract can be increased, and the digestibility of various nutritional ingredients in the feed can be reduced. Similar to soybean meal and other miscellaneous meal, the guar meal also has certain nutritional defects, such as trypsin inhibitor and the like, which influence the utilization rate of protein in animal bodies to a certain extent, and the guar meal has special taste, so that the animal directly has poor taste, and therefore, the method solves the nutritional defects of the guar meal, reduces the toxicity and increases the palatability, and is a hotspot of current research.
At present, the demand of bean pulp is increasing day by day, domestic protein feed resources are insufficient, the import amount of the bean pulp is increasing year by year, and meanwhile, as the European Union issues a ban of forbidding use of feed additives from animal sources and the animal diseases are inundated all over the world, the biological safety problem of the feed is receiving wide attention, which all prompts nutriologists to search for alternative plant protein. Under the pressure that the demand of the soybean meal is increasing day by day, domestic protein feed resources are insufficient, and the import amount of the soybean meal is increased year by year, the guar meal which is a novel protein feed is researched and developed, harmful substances or anti-nutritional factors in the protein feed are removed, the nutritional value of the protein feed is improved, the protein feed becomes high-quality and economic functional protein, and the protein feed has important significance and value for promoting the development of the feed industry and the animal husbandry.
Disclosure of Invention
Aiming at the needs of the prior art, the invention provides a bacillus licheniformis for improving the crude protein content of guar meal and application thereof.
The purpose of the invention is realized by the following technical scheme:
bacillus licheniformis for improving the yield of crude protein in fermentation of guar meal, wherein the preservation number of the Bacillus licheniformis (Bacillus lincheniformis) is CGMCC No. 14185; the preservation unit is as follows: china general microbiological culture Collection center; the address of the preservation unit is as follows: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North; the preservation date is as follows: 24/05/2017.
The invention also provides application of the bacillus licheniformis in guar meal fermentation.
The invention further provides a fermentation method for improving the protein yield in the crude fermentation of the guar meal, which comprises the step of mixing the bacterial liquid containing the bacillus licheniformis and the fermentation culture medium containing the guar meal and then fermenting.
In one embodiment of the invention, the fermentation medium consists of guar meal and bran, wherein the mass ratio of the guar meal to the bran is 9-1: 1.
in one embodiment according to the present invention, the mass ratio of guar meal to bran in the fermentation medium is 1: 1.
in one embodiment of the invention, the ratio of the bacteria liquid to the culture medium is 1-4: 2.
in one embodiment of the invention, the ratio of the bacteria liquid to the culture medium is 5: 4.
the invention further provides a fermentation medium suitable for the bacillus licheniformis, which comprises guar meal and bran, wherein the mass ratio of the guar meal to the bran is (9-1): 1.
in one embodiment according to the present invention, the mass ratio of guar meal to bran is 1: 1.
compared with the prior art, the invention has at least the following advantages:
the invention provides bacillus licheniformis for fermenting guar meal, which can obviously improve the content of crude protein after fermenting the guar meal, and also provides a fermentation method and a corresponding fermentation culture medium suitable for the bacillus licheniformis. The fermentation method and the corresponding fermentation culture medium can obviously improve the yield of crude protein in the fermentation product, and are beneficial to improving the quality and the nutritive value of the feed.
Drawings
FIG. 1 is a bar graph of crude protein content after fermentation of selected Bacillus licheniformis bacteria with different yeast strains on the same medium; wherein 1 is blank control, 2 is yeast 1355, 3 is yeast 1421, 4 is yeast 31015, 5 is yeast 31923, 6 is yeast 32236, and 7 is selected strain;
FIG. 2 shows the results of fermentation of screened Bacillus licheniformis bacteria on media of different formulation ratios; wherein, 1 is the guar meal: the culture medium is prepared by mixing bran 18:2, wherein 2 is prepared from guar meal: the culture medium is prepared by mixing bran 16:4 in a mass ratio, and 3 is prepared by mixing guar meal: the culture medium is prepared from bran 14:6 by mass ratio, and 4 is prepared from guar meal: the culture medium is prepared from bran 12:8 in a mass ratio, and 5 is prepared from guar meal: a culture medium prepared by the weight ratio of 10:10 of bran;
FIG. 3 shows the results of fermentation with different volumes of bacterial liquid added to the same fermentation medium; wherein, 1 is adding 10ml of bacterial liquid, 2 is adding 15ml of bacterial liquid, 3 is adding 20ml of bacterial liquid, 4 is adding 25ml of bacterial liquid, 5 is adding 30ml of bacterial liquid, and 6 is adding 40ml of bacterial liquid.
Detailed Description
The present invention will be further described with reference to the following drawings and examples, which are illustrative only and not intended to be limiting, and the scope of the present invention is not limited thereby.
Reagents and materials:
the reagents and materials used in the present invention are commercially available except for the self-screened Bacillus licheniformis. The preservation number of the bacillus licheniformis (Bacillus lincheniformis) obtained by self-screening is CGMCC No. 14185; the preservation unit is as follows: china general microbiological culture Collection center; the address of the preservation unit is as follows: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North; the preservation date is as follows: 24/05/2017.
The formulation of the medium used in the screening, isolation and culture experiments of the present invention is as follows:
enrichment culture medium: 3g/L of beef extract, 10g/L of peptone, 5g/L of sodium chloride and 7.0 of pH.
Separating a culture medium: 3g/L of beef extract, 10g/L of peptone, 5g/L of sodium chloride, 15g/L of agar and 7.0 of pH.
Yeast culture medium: 10g/L glucose, 10g/L peptone and 5g/L yeast.
The preparation and use of the above culture medium can be performed according to the molecular cloning experimental instruction.
Example 1 Strain screening
Collecting a soil sample from a certain field in Shandong Jinnan, inoculating the soil sample into an enrichment culture solution, culturing for 2-3 days at 37 ℃ and 200rpm, then transferring into the enrichment culture solution, culturing for one day at 37 ℃ and 200rpm, then transferring into a separation culture medium, and standing and culturing in a constant-temperature incubator at 37 ℃. The strain obtained by separation is measured for fermentation characteristics of the strain on the guar meal, and finally the strain with relatively high crude protein yield is obtained.
Example 2 species identification
1. Morphological characteristics: gram staining is positive, the thallus is single and rod-shaped, and generally single generation has flagella. Spores are generated, and the spores are grown. The bacterial colony is crenate, the surface is not smooth and is white.
2. Physiological and biochemical characteristics: through physiological and biochemical characteristic identification, the bacterium can utilize glucose, xylose, arabinose, mannitol and the like to produce acid, and can also utilize citrate and sodium pyruvate to decompose casein and can not decompose casein.
3.16S rRNA Gene sequence homology analysis
The strain obtained by screening was used as a template, and PCR amplification was carried out using bacterial primers 27f and 1492r purchased from Shanghai Biotech Co., Ltd. The 27f primer sequence is as follows: 15 '-AGAGTTTGATCCTGGCTCCAG-3'; the 1492r primer sequence is as follows: 25 '-GGTTACCTTGTTACGACTT-3' of SEQ ID NO. The 16sRNA sequence was obtained as SEQ ID NO 3.
Through the sequence comparison with NCBI database, the similarity of the sequence and the Bacillus licheniformis 16S sequence is 100%, and the strain can be judged to be the Bacillus licheniformis (Bacillus licheniformis) by combining the morphological, physiological and biochemical characteristics.
Example 3 determination of protein content
Yeasts are used in the prior art for the fermentative production of crude proteins, and therefore the screened Bacillus licheniformis strains are compared in this example with several commonly used yeast strains.
Culturing the screened bacillus licheniformis strain and different yeast strains in an enrichment culture medium and a yeast culture medium respectively for 15h, centrifuging, adding 0.1mM PBS (pH7.0) to prepare a strain suspension, adjusting OD to make the OD of all the strains the same, adding 20mL of the strain solution into a sterilized fermentation culture medium (10g of guar meal and 10g of bran), stirring uniformly, culturing in incubators at 37 ℃ and 30 ℃ for 48h respectively, drying at 80 ℃, taking 20g of the ground strain, and sending to a spectronier company to measure the content of crude protein (adding a blank culture medium as a control). (detection basis: GB/T6432-1994) the results are shown in FIG. 1, and the content of crude protein in the fermentation product of the Bacillus licheniformis strain screened by the invention is the highest under the same fermentation conditions.
Example 4 comparison of fermentation results of the strains in different media ratios
Culturing the screened bacillus licheniformis strain in an enrichment culture medium for 15h, centrifuging, adding 0.1mM PBS (pH7.0) to prepare a bacterial suspension, adjusting OD to be the same, then respectively adding 20mL of bacterial liquid into sterilized fermentation culture media (guar meal and bran) with different proportions, stirring uniformly, culturing in a 37 ℃ incubator for 48h, drying at 80 ℃, grinding 20g, and then sending to a SpermanizLenzi company to measure the content of crude protein, wherein the measurement result is shown in figure 2. When the mass ratio of the guar meal to the bran is 16: when 4, the content of the produced protein is the highest, and the mass ratio of the guar bean pulp to the bran is 10:10 times. Considering the cost of raw materials, the mass ratio of the finally adopted guar meal to the bran is 10: 10.
Example 5 comparison of fermentation with different volumes of the bacterial liquid of the strain
Culturing the strain in an enrichment medium for 15h, centrifuging, adding 0.1mM PBS (pH7.0) to prepare a strain suspension, adjusting OD to be the same, respectively taking 10, 15, 20, 25, 30 and 40mL of strain liquid into 20g of sterilized fermentation medium (10g of guar meal and 10g of bran), uniformly stirring, culturing in an incubator at 37 ℃ for 48h, drying at 80 ℃, taking 20g of ground strain liquid, sending to Banni company for measuring the content of crude protein, wherein the measurement result is shown in figure 3, and when 25mL of strain liquid is added, namely the mass ratio of the volume of the strain liquid to the fermentation medium is 5: the content of the produced protein is highest when the yield is 4 hours.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are also included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (6)
1. The bacillus licheniformis for improving the yield of crude protein in the fermentation of guar meal is characterized in that the preservation number of the bacillus licheniformis (Bacillus lincheniformis) is CGMCC No. 14185; the preservation unit is as follows: china general microbiological culture Collection center; the address of the preservation unit is as follows: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North; the preservation date is as follows: 24/05/2017.
2. A fermentation method for increasing the protein yield in the crude fermentation of guar meal, which is characterized in that the fermentation method comprises the step of mixing a bacterial liquid containing the bacillus licheniformis of claim 1 with a fermentation culture medium containing the guar meal and then carrying out fermentation.
3. The fermentation method according to claim 2, wherein the fermentation medium consists of guar meal and bran, wherein the mass ratio of the guar meal to the bran is (9-1): 1.
4. the fermentation process of claim 3, wherein the mass ratio of guar meal to bran in the fermentation medium is 1: 1.
5. the fermentation method according to claim 2 or 3, wherein the ratio of the bacterial liquid to the culture medium is (1-4): 2.
6. the fermentation method according to claim 5, wherein the ratio of the bacterial liquid to the culture medium is 5: 4.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710695330.0A CN108330078B (en) | 2017-08-14 | 2017-08-14 | Bacillus licheniformis for improving crude protein yield in guar meal fermentation and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710695330.0A CN108330078B (en) | 2017-08-14 | 2017-08-14 | Bacillus licheniformis for improving crude protein yield in guar meal fermentation and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108330078A CN108330078A (en) | 2018-07-27 |
| CN108330078B true CN108330078B (en) | 2021-10-08 |
Family
ID=62922661
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710695330.0A Active CN108330078B (en) | 2017-08-14 | 2017-08-14 | Bacillus licheniformis for improving crude protein yield in guar meal fermentation and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108330078B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103911327A (en) * | 2014-03-28 | 2014-07-09 | 内蒙古和美科盛生物技术有限公司 | Bacillus licheniformis probiotic preparation and preparation method thereof |
| CN105754888A (en) * | 2015-12-04 | 2016-07-13 | 中国农业科学院农业环境与可持续发展研究所 | Bacillus licheniformis and microbial agent and their application in fermentation bed culture |
| CN105795099A (en) * | 2015-01-03 | 2016-07-27 | 山东和实集团有限公司 | Preparation of new guar meal animal feed additive by mixed microbial solid fermentation |
| CN106520642A (en) * | 2017-01-12 | 2017-03-22 | 北京瓜尔润科技股份有限公司 | Bacillus amyloliquefaciens and application thereof |
-
2017
- 2017-08-14 CN CN201710695330.0A patent/CN108330078B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103911327A (en) * | 2014-03-28 | 2014-07-09 | 内蒙古和美科盛生物技术有限公司 | Bacillus licheniformis probiotic preparation and preparation method thereof |
| CN105795099A (en) * | 2015-01-03 | 2016-07-27 | 山东和实集团有限公司 | Preparation of new guar meal animal feed additive by mixed microbial solid fermentation |
| CN105754888A (en) * | 2015-12-04 | 2016-07-13 | 中国农业科学院农业环境与可持续发展研究所 | Bacillus licheniformis and microbial agent and their application in fermentation bed culture |
| CN106520642A (en) * | 2017-01-12 | 2017-03-22 | 北京瓜尔润科技股份有限公司 | Bacillus amyloliquefaciens and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 混菌发酵马铃薯淀粉废渣与汁水产单细胞蛋白的研究;石茂萍;《中国优秀硕士学位论文全文数据库 工程科技I辑》;20160215;第40页 * |
| 石茂萍.混菌发酵马铃薯淀粉废渣与汁水产单细胞蛋白的研究.《中国优秀硕士学位论文全文数据库 工程科技I辑》.2016,第40页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108330078A (en) | 2018-07-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108208316B (en) | Siamese bacillus-containing feed | |
| Jeevana Lakshmi et al. | Efficient degradation of feather by keratinase producing Bacillus sp. | |
| CN110218688B (en) | Bacillus marinus LY-23, microbial inoculum and application thereof and product using same | |
| CN106520642B (en) | Bacillus amyloliquefaciens and application thereof | |
| CN110591943B (en) | Bacillus subtilis capable of producing complex enzyme, composition and application thereof | |
| EP3308652B1 (en) | Method for concentrating protein in grain powder | |
| CN107535671B (en) | Microbial fermentation yellow wine lees feed for improving rumen protein utilization rate and preparation method thereof | |
| CN102823731A (en) | Method for preparing small feed composite peptide | |
| JP2019054786A (en) | Bacillus subtilis strain for preparing germ fermentation feather powder and use thereof | |
| CN104222493A (en) | Compound probiotic peptide as well as preparation method and application thereof | |
| KR20120068096A (en) | Microorganism having zearalenone decomposing activity, method for degrading zearalenone using thereof and feed additives comprising the same | |
| CN117603889B (en) | Bacillus subtilis for producing acid protease for feed and application thereof | |
| CN105733999A (en) | Bacillus subtilis FJ-3-16 and application thereof | |
| KR101444196B1 (en) | Novel microorganism Bacillus coagulans KM-1, and manufacturing method of fermented soybean meal and fish feed using the same | |
| CN108338257A (en) | A kind of microbial fermentation zein fiber and its fermentation process and application | |
| CN112980740A (en) | Bacillus licheniformis and application thereof | |
| CN109423466A (en) | A kind of composite fermentation microbial inoculum and its application | |
| CN104286383A (en) | Tea seed meal detoxifying method | |
| CN116622580A (en) | Pediococcus pentosaceus PKC10 and application thereof | |
| CN108330078B (en) | Bacillus licheniformis for improving crude protein yield in guar meal fermentation and application thereof | |
| CN116144528A (en) | Protease-producing lactobacillus reuteri WBT0063 and application thereof in fermented soybean meal and feed | |
| WO2015128366A2 (en) | A method for improving maize digestibility in bovine animals | |
| CN116784417A (en) | High-organic acid fermented palm meal and preparation process thereof | |
| CN103865865B (en) | A kind of sea cucumber enteron aisle produces Sumizyme MP bacterial strain and application thereof | |
| CN108004145B (en) | Black fungus wall breaking method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A Lichen Bacillus spp. for Increasing Crude Protein Yield in Melon Soybean Meal Fermentation and Its Application Effective date of registration: 20221122 Granted publication date: 20211008 Pledgee: Haidian Beijing science and technology enterprise financing Company limited by guarantee Pledgor: BEIJING GUAERRUN TECHNOLOGY CO.,LTD. Registration number: Y2022980022924 |
|
| PC01 | Cancellation of the registration of the contract for pledge of patent right | ||
| PC01 | Cancellation of the registration of the contract for pledge of patent right |
Date of cancellation: 20231109 Granted publication date: 20211008 Pledgee: Haidian Beijing science and technology enterprise financing Company limited by guarantee Pledgor: BEIJING GUAERRUN TECHNOLOGY CO.,LTD. Registration number: Y2022980022924 |