CN115161248B - 一种以海带酶解粉为主要基质的多菌种联合发酵生产植物乳杆菌的方法 - Google Patents
一种以海带酶解粉为主要基质的多菌种联合发酵生产植物乳杆菌的方法 Download PDFInfo
- Publication number
- CN115161248B CN115161248B CN202211027188.XA CN202211027188A CN115161248B CN 115161248 B CN115161248 B CN 115161248B CN 202211027188 A CN202211027188 A CN 202211027188A CN 115161248 B CN115161248 B CN 115161248B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus plantarum
- kelp
- fermentation
- bacillus licheniformis
- culture medium
- 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
- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 38
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 38
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 38
- 241000512259 Ascophyllum nodosum Species 0.000 title claims abstract description 36
- 238000000855 fermentation Methods 0.000 title claims abstract description 31
- 230000004151 fermentation Effects 0.000 title claims abstract description 31
- 239000000843 powder Substances 0.000 title claims abstract description 26
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 10
- 239000011159 matrix material Substances 0.000 title claims abstract description 7
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 25
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims abstract description 25
- 239000001963 growth medium Substances 0.000 claims abstract description 25
- 241000194108 Bacillus licheniformis Species 0.000 claims abstract description 20
- 238000002360 preparation method Methods 0.000 claims abstract description 9
- 239000004382 Amylase Substances 0.000 claims abstract description 5
- 102000013142 Amylases Human genes 0.000 claims abstract description 5
- 108010065511 Amylases Proteins 0.000 claims abstract description 5
- 108091005804 Peptidases Proteins 0.000 claims abstract description 5
- 239000004365 Protease Substances 0.000 claims abstract description 5
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 5
- 235000019418 amylase Nutrition 0.000 claims abstract description 5
- 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 claims description 7
- 239000008103 glucose Substances 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 102000004190 Enzymes Human genes 0.000 claims description 5
- 108090000790 Enzymes Proteins 0.000 claims description 5
- 240000008042 Zea mays Species 0.000 claims description 5
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 5
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 5
- 235000005822 corn Nutrition 0.000 claims description 5
- 229940088598 enzyme Drugs 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 5
- 235000012054 meals Nutrition 0.000 claims description 5
- 150000001875 compounds Chemical class 0.000 claims description 4
- 235000013312 flour Nutrition 0.000 claims description 4
- 238000000034 method Methods 0.000 claims description 4
- 235000013311 vegetables Nutrition 0.000 claims description 4
- 235000015099 wheat brans Nutrition 0.000 claims description 4
- -1 pectase Proteins 0.000 claims description 3
- SFAYBQDGCKZKMH-UHFFFAOYSA-N BNCC Chemical compound BNCC SFAYBQDGCKZKMH-UHFFFAOYSA-N 0.000 claims description 2
- 108010059892 Cellulase Proteins 0.000 claims description 2
- 229940106157 cellulase Drugs 0.000 claims description 2
- 239000007921 spray Substances 0.000 claims description 2
- 238000005507 spraying Methods 0.000 claims description 2
- 238000009630 liquid culture Methods 0.000 abstract description 6
- 241000894006 Bacteria Species 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 230000000813 microbial effect Effects 0.000 abstract description 4
- 235000015097 nutrients Nutrition 0.000 abstract description 4
- 230000000529 probiotic effect Effects 0.000 abstract description 3
- 241000235342 Saccharomycetes Species 0.000 abstract description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 2
- 230000007062 hydrolysis Effects 0.000 abstract description 2
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 2
- 239000001301 oxygen Substances 0.000 abstract description 2
- 229910052760 oxygen Inorganic materials 0.000 abstract description 2
- 239000002609 medium Substances 0.000 description 9
- 238000011081 inoculation Methods 0.000 description 7
- 238000011282 treatment Methods 0.000 description 4
- 238000012258 culturing Methods 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 239000002054 inoculum Substances 0.000 description 3
- 241000206602 Eukaryota Species 0.000 description 2
- 229920000855 Fucoidan Polymers 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 229920000615 alginic acid Polymers 0.000 description 2
- 235000010443 alginic acid Nutrition 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 235000008429 bread Nutrition 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 230000033458 reproduction Effects 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- DBTMGCOVALSLOR-DEVYUCJPSA-N (2s,3r,4s,5r,6r)-4-[(2s,3r,4s,5r,6r)-3,5-dihydroxy-6-(hydroxymethyl)-4-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy-6-(hydroxymethyl)oxane-2,3,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](CO)O[C@H](O)[C@@H]2O)O)O[C@H](CO)[C@H]1O DBTMGCOVALSLOR-DEVYUCJPSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 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
- 208000036649 Dysbacteriosis Diseases 0.000 description 1
- 208000027244 Dysbiosis Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- 229920001543 Laminarin Polymers 0.000 description 1
- 239000005717 Laminarin Substances 0.000 description 1
- UPYKUZBSLRQECL-UKMVMLAPSA-N Lycopene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1C(=C)CCCC1(C)C)C=CC=C(/C)C=CC2C(=C)CCCC2(C)C UPYKUZBSLRQECL-UKMVMLAPSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 1
- 241000199919 Phaeophyceae Species 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 241000015177 Saccharina japonica Species 0.000 description 1
- 241000235343 Saccharomycetales Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 229920001284 acidic polysaccharide Polymers 0.000 description 1
- 150000004805 acidic polysaccharides Chemical class 0.000 description 1
- IAJILQKETJEXLJ-RSJOWCBRSA-N aldehydo-D-galacturonic acid Chemical compound O=C[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)C(O)=O IAJILQKETJEXLJ-RSJOWCBRSA-N 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 238000000889 atomisation Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003833 bile salt Substances 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- KLOIYEQEVSIOOO-UHFFFAOYSA-N carbocromen Chemical compound CC1=C(CCN(CC)CC)C(=O)OC2=CC(OCC(=O)OCC)=CC=C21 KLOIYEQEVSIOOO-UHFFFAOYSA-N 0.000 description 1
- 150000001746 carotenes Chemical class 0.000 description 1
- 235000005473 carotenes Nutrition 0.000 description 1
- 230000034303 cell budding Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 229940079919 digestives enzyme preparation Drugs 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 230000007140 dysbiosis Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 235000021107 fermented food Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 235000003642 hunger Nutrition 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000011177 media preparation Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 150000004804 polysaccharides Chemical class 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229960003080 taurine Drugs 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- NCYCYZXNIZJOKI-UHFFFAOYSA-N vitamin A aldehyde Natural products O=CC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C NCYCYZXNIZJOKI-UHFFFAOYSA-N 0.000 description 1
- 239000007222 ypd medium Substances 0.000 description 1
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
-
- 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
-
- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
-
- 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
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- 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
- C12R2001/10—Bacillus licheniformis
-
- 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
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
-
- 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
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/85—Saccharomyces
- C12R2001/865—Saccharomyces cerevisiae
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Virology (AREA)
- General Health & Medical Sciences (AREA)
- Mycology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Animal Husbandry (AREA)
- Physiology (AREA)
- Food Science & Technology (AREA)
- Botany (AREA)
- Molecular Biology (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明公开了一种以海带酶解粉为主要基质的多菌种联合发酵生产植物乳杆菌的方法,属于微生物发酵技术领域。本发明利用酿酒酵母、地衣芽孢杆菌和植物乳杆菌进行联合发酵,可生产高活性饲料用微生态制剂产品。在液态培养基中,先通过酵母菌的生长消耗掉其中的氧气,创造厌氧环境,然后通过地衣芽孢杆菌生长分泌蛋白酶、淀粉酶等,水解培养基中养分可供植物乳杆菌快速生长。三种菌的联合培养充分利用了海带酶解物等培养基中的各种营养成分,将其转化成高益生活性的植物乳杆菌菌体。
Description
技术领域
本发明属于微生物发酵技术领域,尤其涉及一种以海带酶解粉为主要基质的多菌种联合发酵生产植物乳杆菌的方法。
背景技术
海带(Laminariajaponica)是多年生大型食用藻类,含有丰富的褐藻胶、岩藻糖胶、褐藻淀粉等海带多糖,并含有酸性聚糖类物质、岩藻半乳多糖硫酸酯、大叶藻素、半乳糖醛酸、昆布氨酸、牛磺酸、双歧因子等多种活性成分。据测定,每100g海带中含8g蛋白质、0.1g脂肪、0.57mg胡萝卜素、0.09mgVB、0.36mgVB1、1.6mg烟酸、1.177mg钙、1.50mg铁,2.16mg磷,还含有丰富的褐藻酸、纤维素、甘露醇以及多种微量元素。海带酶解干粉是通过多种酶制剂将海带原浆进行降解,充分释放里面的活性成分,然后喷雾干燥处理成干粉。海带酶解干粉里面含有多种氨基酸、维生素、多糖和微量元素,非常适合作为微生物发酵培养基使用。利用微生物发酵,将低值的海带酶解粉转化为高活性微生态制剂产品,可以大幅度提高其附加值。
酿酒酵母(Saccharomyces cerevisiae),又称面包酵母或者出芽酵母。酿酒酵母是与人类关系最广泛的一种酵母,用于制作面包和馒头等食品及酿酒。酿酒酵母的细胞为球形或者卵形,直径5-10μm。其繁殖的方法为出芽生殖。酿酒酵母与同为真核生物的动物和植物细胞具有很多相同的结构,又容易培养,酵母被用作研究真核生物的模式生物。酿酒酵母被认为是最具潜力的大规模生产菌种。
地衣芽孢杆菌(Bacillus licheniformis)是一种革兰氏染色阳性菌,能够分泌蛋白酶、淀粉酶等多种酶,酶分泌的最适温度为37℃该细菌可调整菌群失调达到治疗目的,可促使机体产生抗菌活性物质、杀灭致病菌。能产生抗活性物质抑制致病菌的生长繁殖。
植物乳杆菌(Lactobacillusplantarum)是一种在自然界广泛存在的革兰氏阳性菌,普遍分布在各类发酵食品中,同时也是肠道内重要的益生微生物菌群之一。由于植物乳杆菌具有优良的耐受强酸、胆盐、高渗透压、极端温度、饥饿和氧化等胁迫的能力,使得其在工业生产、医疗保健及食品加工等领域被广泛应用。
发明内容
本发明的目的之一在于提供一种以海带酶解粉为主要基质的多菌种联合发酵生产植物乳杆菌的方法,所述方法是以酿酒酵母、地衣芽孢杆菌和植物乳杆菌作为发酵菌种,在含有海带酶解物干粉的培养基中进行发酵。
优选地,所述酿酒酵母、地衣芽孢杆菌和植物乳杆菌分别为酿酒酵母BNCC192858、地衣芽孢杆菌SDHY-2101和植物乳杆菌BNCC 138404。
更优选地,所述酿酒酵母BNCC192858、地衣芽孢杆菌SDHY-2101和植物乳杆菌BNCC138404的种子液的体积比为1:2:2或2:2:2。
更优选地,所述含有海带酶解物干粉的培养基为:海带酶解物干粉10%-19wt%、菜粕8wt%、葡萄糖3wt%、小麦麸皮8wt%、玉米粉5wt%、余量为水。
优选地,所述发酵过程为恒温发酵,温度为30℃,发酵时间为72h。
优选地,所述发酵过程分为两个阶段,第一个阶段为30℃培养24h,第二个阶段为37℃继续培养48h。
本发明的目的之二在于提供上述以海带酶解粉为主要基质的多菌种联合发酵生产植物乳杆菌的方法在生产植物乳杆菌中的应用。
与现有技术相比,本发明具有如下有益效果:
本发明利用酿酒酵母、地衣芽孢杆菌和植物乳杆菌进行联合发酵,可生产高活性饲料用微生态制剂产品。在液态培养基中,先通过酵母菌的生长消耗掉其中的氧气,创造厌氧环境,然后通过地衣芽孢杆菌生长分泌蛋白酶、淀粉酶等,水解培养基中养分可供植物乳杆菌快速生长。三种菌的联合培养充分利用了海带酶解物等培养基中的各种营养成分,将其转化成高益生活性的植物乳杆菌菌体。
具体实施方式
实施例1酿酒酵母BNCC192858种子液的制备
YPD培养基配制:酵母提取物10.0g、蛋白胨20.0g、葡萄糖20.0g,蒸馏水溶解,定容至1000mL,121℃高压灭菌20min。
取酿酒酵母BNCC192858单菌落接种于10mLYPD液体培养基,30℃、220r/min振荡培养12h。吸取100μL,转接100mLYPD培养基,继续28℃、220r/min振荡培养18-24h。
实施例2地衣芽孢杆菌SDHY-2101种子液的制备
培养基配制:葡萄糖10.0g、玉米粉12.0g、酵母粉3.0g、海带酶解干粉5.0g,定容至1000mL,121℃高压灭菌20min。
地衣芽孢杆菌SDHY-2101由本公司实验室保存。挑取地衣芽孢杆菌单菌落接种10mL培养基,37℃、220r/min振荡培养12h。吸取100μl菌液转接100mL培养基中,继续37℃、220r/min振荡培养24h。
实施例3植物乳杆菌BNCC 138404种子液制备。
MRS培养基配制:蛋白胨10.0g、牛肉膏10.0g、酵母膏5.0g、柠檬酸氢二铵2.0g、葡萄糖20.0g、吐温-801.0mL、乙酸钠5.0g、磷酸氢二钾2.0g、MgSO4·7H2O 0.50g、MnSO4·H2O0.25g,蒸馏水溶解,定容至1000mL,pH在6.2-6.6之间。121℃高压灭菌20min。
植物乳杆菌菌种(BNCC138404)购北京北纳创联生物技术研究院。挑取植物乳杆菌单菌落,接种10mLMRS液体培养基,37℃静止培养24h。吸取1.0mL,转接至100mLMRS培养基,继续37℃静止培养24h。
实施例4海带酶解物干粉的制备
鲜海带以1:1-1:1.5的比例加水粉碎后加入0.1%-0.5%的复合酶(复合酶中纤维素酶、果胶酶、蛋白酶和淀粉酶的质量百分数分别为60%、30%、5%和5%),40-60℃酶解20-24h,获得海带酶解液,海带酶解液经喷雾干燥机(设定送风频率15-22HZ、雾化频率225-235HZ、引风频率35-45HZ、进风温度170-180℃、出风温度75-85℃)进行喷粉。
实施例5发酵培养基的制备
培养基主要成分为:海带酶解物干粉10%-19wt%、菜粕8wt%、葡萄糖3wt%、小麦麸皮8wt%、玉米粉5wt%,具体配方见表1,pH值自然。500mL三角瓶中装50g,封口膜密闭封口,121℃高压灭菌20min。
表1发酵培养基配方
组分 | 培养基1 | 培养基2 | 培养基3 | 培养基4 |
海带酶解干粉 | 10% | 13% | 16% | 19% |
菜粕 | 8% | 8% | 8% | 8% |
玉米粉 | 5% | 5% | 5% | 5% |
葡萄糖 | 3% | 3% | 3% | 3% |
小麦麸皮 | 8% | 8% | 8% | 8% |
KH2PO4 | 2% | 2% | 2% | 2% |
NaCl | 1% | 1% | 1% | 1% |
蒸馏水 | 63% | 60% | 57% | 54% |
实施例6恒温发酵工艺
接种剂量组合:将培养好的酿酒酵母、地衣芽孢杆菌和植物乳杆菌的种子液按照体积比1:2:2(接种组合1)和2:2:2(接种组合2)的比例(见表2),在超净工作台中接种(接种量为千分之一),接种后摇匀,密封瓶口。每种培养基做3个平行处理,置于30℃培养箱中培养72h。
表2各菌株在发酵培养基中的接种比例
实施例7变温发酵工艺
接种剂量组合:将培养好的酿酒酵母、地衣芽孢杆菌和植物乳杆菌的种子液按照体积比1:2:2(接种组合1)和2:2:2(接种组合2)的比例(见表2),在超净工作台中接种,接种总量同实施例6,接种后摇匀,密封瓶口。每种培养基做3个平行处理,先置于30℃培养箱中培养24h,调整培养温度为37℃继续培养48h。
实施例8培养物pH值及活菌计数
发酵完成后,测定培养物pH值和植物乳杆菌的活菌数(见表3、4)。植物乳杆菌的活菌计数采用GB/T 4789.35的方法进行。从结果可知,第2种接种组合中,第3种培养基的发酵效果最好,pH值最低,活菌数量最多。变温培养的活菌数高于恒温培养的活菌数。
表3恒温培养物的pH值和植物乳杆菌活菌数
表4变温培养物的pH值和植物乳杆菌活菌数
对比例1
配制第3种培养基(参见实施例5中的表1),灭菌后,等体积比例(2:2)接种酿酒酵母和植物乳杆菌或者地衣芽孢杆菌和植物乳杆菌,接种总量同实施例6,每个接种剂量设置3个重复,然后按照变温培养方案进行发酵培养,测定pH值和植物乳杆菌的活菌数(表5)。
表5
以上所述的实施例仅是对本发明的优选方式进行描述,并非对本发明的范围进行限定,在不脱离本发明设计精神的前提下,本领域普通技术人员对本发明的技术方案做出的各种变形和改进,均应落入本发明权利要求书确定的保护范围内。
Claims (3)
1.一种以海带酶解粉为主要基质的多菌种联合发酵生产植物乳杆菌的方法,其特征在于,所述方法是以酿酒酵母、地衣芽孢杆菌和植物乳杆菌作为发酵菌种,在含有海带酶解物干粉的培养基中进行发酵;
所述酿酒酵母、地衣芽孢杆菌和植物乳杆菌分别为酿酒酵母BNCC192858、地衣芽孢杆菌SDHY-2101和植物乳杆菌BNCC 138404;
所述酿酒酵母BNCC192858、地衣芽孢杆菌SDHY-2101和植物乳杆菌BNCC 138404的种子液的体积比为1:2:2或2:2:2;
所述含有海带酶解物干粉的培养基为:海带酶解物干粉10%-19wt%、菜粕8wt%、葡萄糖3wt%、小麦麸皮8wt%、玉米粉5wt%、余量为水;
所述发酵过程为恒温发酵,温度为30℃,发酵时间为72h;
所述海带酶解物干粉的制备:鲜海带以1:1-1:1.5的比例加水粉碎后加入0.1%-0.5%的复合酶,其中复合酶中纤维素酶、果胶酶、蛋白酶和淀粉酶的质量百分数分别为60%、30%、5%和5%,40-60℃酶解20-24h,获得海带酶解液,海带酶解液经喷雾干燥机进行喷粉。
2.根据权利要求1所述的方法,其特征在于,所述发酵过程分为两个阶段,第一个阶段为30℃培养24h,第二个阶段为37℃继续培养48h。
3.权利要求1-2中任一项所述的方法在生产植物乳杆菌中的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202211027188.XA CN115161248B (zh) | 2022-08-25 | 2022-08-25 | 一种以海带酶解粉为主要基质的多菌种联合发酵生产植物乳杆菌的方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202211027188.XA CN115161248B (zh) | 2022-08-25 | 2022-08-25 | 一种以海带酶解粉为主要基质的多菌种联合发酵生产植物乳杆菌的方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN115161248A CN115161248A (zh) | 2022-10-11 |
CN115161248B true CN115161248B (zh) | 2023-11-21 |
Family
ID=83481229
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202211027188.XA Active CN115161248B (zh) | 2022-08-25 | 2022-08-25 | 一种以海带酶解粉为主要基质的多菌种联合发酵生产植物乳杆菌的方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN115161248B (zh) |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114214247A (zh) * | 2021-12-27 | 2022-03-22 | 威海市世代海洋生物科技股份有限公司 | 一株地衣芽孢杆菌及其应用 |
CN114532460A (zh) * | 2022-01-20 | 2022-05-27 | 大连民族大学 | 混合菌在海带、马尾藻微生物发酵饲料中的应用及其制备方法 |
CN114891663A (zh) * | 2022-04-13 | 2022-08-12 | 山东筱萤生物科技有限公司 | 植物乳杆菌lp1406及分离培养方法 |
CN114903154A (zh) * | 2022-04-13 | 2022-08-16 | 山东筱萤生物科技有限公司 | 一种基于酵母-乳酸菌分步发酵获得降血糖型发酵海藻产品的方法 |
-
2022
- 2022-08-25 CN CN202211027188.XA patent/CN115161248B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114214247A (zh) * | 2021-12-27 | 2022-03-22 | 威海市世代海洋生物科技股份有限公司 | 一株地衣芽孢杆菌及其应用 |
CN114532460A (zh) * | 2022-01-20 | 2022-05-27 | 大连民族大学 | 混合菌在海带、马尾藻微生物发酵饲料中的应用及其制备方法 |
CN114891663A (zh) * | 2022-04-13 | 2022-08-12 | 山东筱萤生物科技有限公司 | 植物乳杆菌lp1406及分离培养方法 |
CN114903154A (zh) * | 2022-04-13 | 2022-08-16 | 山东筱萤生物科技有限公司 | 一种基于酵母-乳酸菌分步发酵获得降血糖型发酵海藻产品的方法 |
Non-Patent Citations (1)
Title |
---|
Effect of Microbial Fermentation on the Fishy-Odor Compounds in Kelp (Laminaria japonica);Wenyang Zhu et al.;foods;全文 * |
Also Published As
Publication number | Publication date |
---|---|
CN115161248A (zh) | 2022-10-11 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN103275876B (zh) | 茶源冠突散囊菌菌株及其应用 | |
CN103555598B (zh) | 一种食用酵素复合菌剂及其制备方法 | |
CN102409007B (zh) | 一种芽孢杆菌微生态制剂及其液固结合发酵制备工艺 | |
CN112457999B (zh) | 一种富硒酿酒酵母菌株及其应用 | |
CN103184174B (zh) | 培养基中含腐植酸钠饲料用枯草芽孢杆菌生物制剂的生产 | |
CN103305434A (zh) | 具有益生菌和有机硒双重作用的微生态制剂及其制备方法 | |
CN102805209A (zh) | 一种畜禽用合生素饲料添加剂及其应用 | |
CN101974601A (zh) | 一种r-多糖发酵液的制备及其复合防腐剂的制备方法 | |
CN106922952B (zh) | 一种米曲霉固态发酵制备富多肽蛋白饲料添加剂的方法 | |
CN111642622A (zh) | 一种发酵复合菌和桃金娘发酵提取物的制备及应用 | |
CN111700157A (zh) | 一种提高水产动物免疫力的益生菌饲料添加剂 | |
CN112210502B (zh) | 草酸青霉突变株a2-13及其在制备生淀粉酶制剂与降解生淀粉中的应用 | |
CN103289935A (zh) | 一种复合菌株微生态制剂及其制备方法 | |
CN105265769B (zh) | 一种多功能菌发酵复合酶饲料及其制备方法 | |
CN112280707A (zh) | 一种饲料用益生菌的制备方法 | |
CN109943502B (zh) | 益生菌、发酵剂及在中草药制剂中的应用 | |
CN108354172B (zh) | 一种樟芝蘑菇酵素的制备方法 | |
CN114617202A (zh) | 一种提高鸭抗病力和生产性能的复合微生态制剂、其制备方法与应用 | |
CN103202386B (zh) | 一种干酪乳杆菌发酵液在饲料中的应用 | |
CN109363004A (zh) | 大鳞鲃鱼用发酵型中草药免疫增强剂的制备方法与应用 | |
CN106479899B (zh) | 一种蛹虫草菌株及其在制备虫草素中的应用 | |
CN115161248B (zh) | 一种以海带酶解粉为主要基质的多菌种联合发酵生产植物乳杆菌的方法 | |
CN101244088A (zh) | 一种肉鸡专用的抗病微生物制剂 | |
CN102669432B (zh) | 一种鹅用复合益生菌制剂及其制备方法 | |
KR100396022B1 (ko) | 가축용 생균제 및 가축용 생균제의 제조방법 |
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 | ||
CB03 | Change of inventor or designer information |
Inventor after: Qu Yuankai Inventor after: Li Mingtan Inventor before: Li Mingtan Inventor before: Qu Yuankai |
|
CB03 | Change of inventor or designer information | ||
GR01 | Patent grant | ||
GR01 | Patent grant |