CN108865919A - 一种戊糖乳杆菌cicc6294及其在辣椒发酵中的应用 - Google Patents
一种戊糖乳杆菌cicc6294及其在辣椒发酵中的应用 Download PDFInfo
- Publication number
- CN108865919A CN108865919A CN201710328555.2A CN201710328555A CN108865919A CN 108865919 A CN108865919 A CN 108865919A CN 201710328555 A CN201710328555 A CN 201710328555A CN 108865919 A CN108865919 A CN 108865919A
- Authority
- CN
- China
- Prior art keywords
- capsicum
- cicc
- nitrite
- lactobacillus pentosus
- bacterial strain
- 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.)
- Granted
Links
- 241000186684 Lactobacillus pentosus Species 0.000 title claims abstract description 37
- 235000002566 Capsicum Nutrition 0.000 title claims abstract description 36
- 239000001390 capsicum minimum Substances 0.000 title claims abstract description 36
- 238000000855 fermentation Methods 0.000 title claims abstract description 13
- 230000004151 fermentation Effects 0.000 title claims abstract description 13
- 240000008574 Capsicum frutescens Species 0.000 title 1
- 241000208293 Capsicum Species 0.000 claims abstract description 35
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 claims abstract description 34
- 150000001412 amines Chemical class 0.000 claims abstract description 28
- 230000000035 biogenic effect Effects 0.000 claims abstract description 23
- 230000001580 bacterial effect Effects 0.000 claims abstract description 17
- 230000015556 catabolic process Effects 0.000 claims abstract description 16
- 238000006731 degradation reaction Methods 0.000 claims abstract description 16
- 102000004031 Carboxy-Lyases Human genes 0.000 claims abstract description 8
- 108090000489 Carboxy-Lyases Proteins 0.000 claims abstract description 8
- 150000001413 amino acids Chemical class 0.000 claims abstract description 8
- 235000013305 food Nutrition 0.000 claims abstract description 8
- 241000588724 Escherichia coli Species 0.000 claims abstract description 5
- 241000186779 Listeria monocytogenes Species 0.000 claims abstract description 5
- 241000191967 Staphylococcus aureus Species 0.000 claims abstract description 5
- 241000607142 Salmonella Species 0.000 claims abstract description 3
- 238000009629 microbiological culture Methods 0.000 claims abstract 2
- 235000012055 fruits and vegetables Nutrition 0.000 claims description 8
- 239000012530 fluid Substances 0.000 claims description 2
- 235000015097 nutrients Nutrition 0.000 claims description 2
- 241000192125 Firmicutes Species 0.000 claims 1
- 201000010099 disease Diseases 0.000 claims 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims 1
- GQPLMRYTRLFLPF-UHFFFAOYSA-N nitrous oxide Inorganic materials [O-][N+]#N GQPLMRYTRLFLPF-UHFFFAOYSA-N 0.000 claims 1
- 244000000010 microbial pathogen Species 0.000 abstract description 7
- 239000001963 growth medium Substances 0.000 abstract description 3
- 230000005764 inhibitory process Effects 0.000 abstract description 3
- 244000005700 microbiome Species 0.000 abstract description 3
- 238000009630 liquid culture Methods 0.000 abstract 1
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 22
- 241000894006 Bacteria Species 0.000 description 17
- 235000014655 lactic acid Nutrition 0.000 description 11
- 239000004310 lactic acid Substances 0.000 description 11
- 239000002253 acid Substances 0.000 description 7
- 238000011081 inoculation Methods 0.000 description 7
- 229910002651 NO3 Inorganic materials 0.000 description 6
- 108020004465 16S ribosomal RNA Proteins 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 235000021121 fermented vegetables Nutrition 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 101150080777 pheS gene Proteins 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 150000003839 salts Chemical class 0.000 description 4
- IOVCWXUNBOPUCH-UHFFFAOYSA-N Nitrous acid Chemical compound ON=O IOVCWXUNBOPUCH-UHFFFAOYSA-N 0.000 description 3
- 230000003385 bacteriostatic effect Effects 0.000 description 3
- 230000000593 degrading effect Effects 0.000 description 3
- 230000000877 morphologic effect Effects 0.000 description 3
- 230000002269 spontaneous effect Effects 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 241000186660 Lactobacillus Species 0.000 description 2
- UCTWMZQNUQWSLP-UHFFFAOYSA-N adrenaline Chemical compound CNCC(O)C1=CC=C(O)C(O)=C1 UCTWMZQNUQWSLP-UHFFFAOYSA-N 0.000 description 2
- 230000004087 circulation Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 229940039696 lactobacillus Drugs 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- KIDHWZJUCRJVML-UHFFFAOYSA-N putrescine Chemical compound NCCCCN KIDHWZJUCRJVML-UHFFFAOYSA-N 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 235000013311 vegetables Nutrition 0.000 description 2
- 239000004475 Arginine Substances 0.000 description 1
- 108090000121 Aromatic-L-amino-acid decarboxylases Proteins 0.000 description 1
- 102000003823 Aromatic-L-amino-acid decarboxylases Human genes 0.000 description 1
- 102100035882 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- XUIIKFGFIJCVMT-GFCCVEGCSA-N D-thyroxine Chemical compound IC1=CC(C[C@@H](N)C(O)=O)=CC(I)=C1OC1=CC(I)=C(O)C(I)=C1 XUIIKFGFIJCVMT-GFCCVEGCSA-N 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 206010012289 Dementia Diseases 0.000 description 1
- 101100311938 Dictyostelium discoideum phesA gene Proteins 0.000 description 1
- 101100423325 Dictyostelium discoideum phesB gene Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 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 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- 241000194036 Lactococcus Species 0.000 description 1
- 108010048581 Lysine decarboxylase Proteins 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 102000052812 Ornithine decarboxylases Human genes 0.000 description 1
- 108700005126 Ornithine decarboxylases Proteins 0.000 description 1
- 241000192001 Pediococcus Species 0.000 description 1
- 239000005700 Putrescine Substances 0.000 description 1
- 241000194017 Streptococcus Species 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000002479 acid--base titration Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000007269 microbial metabolism Effects 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 238000011392 neighbor-joining method Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- -1 palpitating speed Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000009938 salting Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000004206 stomach function Effects 0.000 description 1
- 238000005482 strain hardening Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 229940034208 thyroxine Drugs 0.000 description 1
- XUIIKFGFIJCVMT-UHFFFAOYSA-N thyroxine-binding globulin Natural products IC1=CC(CC([NH3+])C([O-])=O)=CC(I)=C1OC1=CC(I)=C(O)C(I)=C1 XUIIKFGFIJCVMT-UHFFFAOYSA-N 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 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
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3571—Microorganisms; Enzymes
-
- 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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- 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
- A23V2400/167—Pentosus
-
- 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
Abstract
本发明涉及食品微生物领域，特别涉及到一种高效快速降解亚硝酸盐，低产生物胺且具有抑制致病微生物的戊糖乳杆菌(Lactobacillus pentosus)CICC 6294及其在新疆特色辣椒发酵中的应用。该菌株保藏于中国普通微生物菌种保藏管理中心，保藏编号为：CGMCC 14016。本发明所述的菌株在改良MRS液体培养基中培养24h，亚硝酸盐降解率能够达到98.8%；培养30h，亚硝酸盐降全部降解。此外，该菌株氨基酸脱羧酶为阴性，且能够抑制大肠埃希氏菌、金黄色葡萄球菌、单增李斯特菌和沙门氏菌生长。将该菌株用于新疆特色辣椒发酵，所得到发酵辣椒中亚硝酸盐的含量0.81mg/kg，生物胺的含量为3.19mg/kg，极大的提高发酵辣椒的食用安全性。
Description
技术领域
本发明涉及食品微生物领域，特别涉及到一种高效快速降解亚硝酸盐，低产生物胺且具有抑制致病微生物的戊糖乳杆菌CICC 6294及其在新疆特色辣椒发酵中的应用。
背景技术
乳酸菌是一类能利用可发酵碳水化合物产生大量乳酸的革兰氏染色阳性细菌的总称。乳酸菌在自然界分布极为广泛，具有丰富的物种多样性，目前报道的乳酸菌超过了20个属，主要包括乳杆菌属，乳球菌属，链球菌属，片球菌属等，但它们具有一些共同的特征，包括革兰氏染色阳性、无运动性或很少有运动性、厌氧或兼性厌氧、以乳酸作为发酵代谢中主要或唯一产物等。乳酸菌发酵能够产生大量的有机酸，具有耐酸和耐盐特性，亚硝酸盐降解能力，抑制致病微生物特性以及低产生物胺特性，广泛应用于果蔬发酵生产中。蔬菜发酵是利用有益微生物的作用，控制一定生产条件对蔬菜进行加工的一种方式。蔬菜发酵属于一种冷加工方式,既能够最大程度的保持蔬菜的营养和色香味，又具有良好的感官品质，深受消费者喜爱。蔬菜发酵体系是一多种微生物相互作用的微生态环境，乳酸菌发挥着重要作用。目前由于果蔬种植过程中化肥使用量加大，果蔬中的硝酸盐残留显著增加。果蔬在腌制发酵过程中在硝酸盐还原菌的作用生成亚硝酸盐，人体摄取过量的亚硝酸盐会引起致癌、抵抗甲状腺素和智障等危害。果蔬在腌制发酵过程中在氨基酸脱羧酶的作用下积累大量的生物胺，一旦摄入过量的生物胺，会导致肾上腺素和胃酸过量分泌、心跳加快、血糖含量增加或血压升高等症状。因此，如何降低发酵果蔬中的亚硝酸盐和生物胺含量是亟待解决的技术问题，开发筛选出快速高效降解亚硝酸盐，低产生物胺的乳酸菌菌株将会成为研究的重点。本发明筛选得到的戊糖乳杆菌 CICC 6294具有快速高效降解亚硝酸盐，低产生物胺和抑制致病微生物的特性，应用于新疆特色辣椒发酵生产中，能够降低产品中亚硝酸盐和生物胺的含量，提高发酵辣椒的食用安全性。
发明内容
本发明的目的是提供一株戊糖乳杆菌CICC 6294，其特征在于快速高效的降解亚硝酸盐，低产生物胺，抑制致病微生物生长。
本发明的另一个目的是提供一种接种上述戊糖乳杆菌CICC 6294的亚硝酸盐和生物胺含量低的新疆特色发酵辣椒。
本发明在于克服现有技术的不足，通过接种能够快速高效的降解亚硝酸盐，低产生物胺，抑制致病微生物生长的戊糖乳杆菌CICC 6294，即可生产出亚硝酸盐和生物胺含量低的新疆特色发酵辣椒。
本发明的具体技术方案如下：
戊糖乳杆菌CICC 6294为实验室从新疆特色辣椒中分离得到的乳酸菌，保藏于中国普通微生物菌种保藏中心，保藏编号为CGMCC 14016。该菌株菌落乳白色，圆形，表面光滑，不透明，边缘整齐。菌体呈短杆状，0.4-0.5 μm × 0.6-1.5 μm，成对或成堆排列，革兰氏染色呈阳性，过氧化氢酶阴性。
戊糖乳杆菌CICC 6294具有较强亚硝酸盐降解能力，在MRS 液体培养基中培养24h，亚硝酸盐降解率能够达到98.8%，培养30 h，亚硝酸盐降解率达到100%，亚硝酸盐降解效率和速率都优于以往报道的菌株。戊糖乳杆菌CICC 6294具有较强的抑制致病微生物的能力，能够抑制大肠埃希氏菌、金黄色葡萄球菌、单增李斯特菌和沙门氏菌四种食品常见污染菌的生长。戊糖乳杆菌CICC 6294氨基酸脱羧酶为阴性，在生长代谢过程中产生生物胺的能力较低。
将戊糖乳杆菌CICC 6294应用于新疆特色辣椒的发酵过程中，所得到的发酵辣椒亚硝酸盐的含量0.81 mg/kg, 生物胺的含量为3.19 mg/kg，降低了发酵蔬菜中残留亚硝酸盐和生物胺，解决了亚硝酸盐和生物胺带来的食品安全问题。此外，接种该菌株，能够缩短辣椒发酵周期，并且发酵辣椒的总酸含量达到2.76%。
附图说明
图1示发明所述戊糖乳杆菌CICC 6294的扫描电镜形态。
图2示发明所述戊糖乳杆菌CICC 6294的16S rRNA 基因和pheS基因系统发育树。
图3示发明所述戊糖乳杆菌CICC 6294亚硝酸盐降解特性。
图4示发明所述接种戊糖乳杆菌CICC 6294发酵辣椒中亚硝酸盐含量。
图5示发明所述接种戊糖乳杆菌CICC 6294发酵辣椒中生物胺含量。
具体实施方案
下面结合实施例对本发明进一步说明，下述实施例是说明性的，不是限定性的，不能以下述实施例来限定本发明的保护范围。
实施例1 菌株CICC 6294的鉴定
本发明中的的菌株CICC 6294是从新疆传统发酵辣椒中分离筛选得到，利用形态学和分子生物学等方法将该乳酸菌鉴定为戊糖乳杆菌。
1、形态学鉴定
参照《常见细菌系统鉴定手册》对CICC 6294进行形态特征和鉴定，并对其进行扫描电镜观察（图1）。MRS培养基，37℃培养24 h，菌落乳白色，圆形，表面光滑，不透明，边缘整齐。菌体呈短杆状，0.4-0.5 μm × 0.6-1.5 μm，成对或成堆排列，革兰氏染色阳性，过氧化氢酶阴性。
2、16S rRNA 基因及pheS基因的PCR 扩增
使用细菌基因组DNA提取试剂盒提取乳酸菌的基因组。利用引27f (5’-AGAGTTTGATCCTGGCTCAG-3’) 和1492r (5’-GGTTACCTTGTTACGACTT-3’) 对16S rRNA基因进行扩增。PCR反应体系为：模板10×PCR buffer 5 μL、dNTP (2.5 mmol/L) 4 μL、模板 2 μL、TaqDNA 聚合酶 1 μL、引物各 1 μL, 补充去离子水至50 μL。反应条件: 94℃ 5 min，94℃ 50s，52℃ 50 s，72℃ 50 s，33个循环，72℃ 7 min。
利用引物21f (5’-CAYCCNGCHCGYGAYATGC-3’)和23r (5’-GGRTGRACCATVCCNGCHCC-3’)对pheS基因序列进行扩增。PCR反应体系同上，反应条件：95℃ 5 min，95℃ 1min，46℃ 2 min 15 s，72℃ 1 min 15 s，3个循环；95℃ 1 min 15 s，46℃ 1 min 15 s,72℃ 1 min 15 s，30个循环；72℃ 7 min。
4、序列分析
将所得的16S rRNA 基因和pheS基因序列用BLAST 软件与NCBI数据库进行相似性比对，运用MEGA 5.0 软件采用Neighbor Joining 方法构建系统发育树。菌株CICC 6294与戊糖乳杆菌处于同一个分支（图2），再结合形态学特征，鉴定为戊糖乳杆菌。其16S rRNA 基因序列登录号为KY694989，pheS基因序列登录号为KY694987。
实施例2 戊糖乳杆菌CICC 6294的特性研究
1、亚硝酸盐降解能力
按照GB 5009.33-2010：《食品中亚硝酸盐与硝酸盐的测定》测定戊糖乳杆菌CICC6294亚硝酸盐降解能力。1%接种量接种到含0.125 mg/ mL NaNO2 的MRS培养基中，37℃培养24 h，亚硝酸盐降解率能够达到98.8%，培养30 h，亚硝酸盐降解率达到100%（图3）。
2、氨基酸脱羧酶试验
微生物代谢过程中，在氨基酸脱羧酶作用下，生成生物胺类物质。筛选氨基酸脱羧酶为阴性的菌株作为发酵剂，会降低果蔬发酵生产中生物胺的含量。氨基酸脱羧酶试验结果表明，戊糖乳杆菌CICC 6294对精氨酸双水解、色氨酸脱羧酶、赖氨酸脱羧酶和鸟氨酸脱羧酶活性均为阴性。
3、抑菌能力
利用滤纸片法测定戊糖乳杆菌CICC 6294对肠炎沙门氏菌、大肠杆菌、金黄色葡萄球菌和单增李斯特菌的抑制程度。戊糖乳杆菌CICC 6294过夜培养物对肠炎沙门氏菌的抑菌直径达到26.0 mm，对大肠杆菌的抑菌直径达到13.3mm，对金黄色葡萄球菌的抑菌直径达到19.4 mm，对单增李斯特菌抑菌直径达到10.3 mm。
实施例3 戊糖乳杆菌CICC 6294在新疆特色辣椒发酵中的应用
选择新鲜的新疆辣椒，清洗，破碎后，加入3% 蔗糖和3% 盐，接种1%戊糖乳杆菌CICC6294（108 CFU/mL），30℃恒温发酵7天，即得到发酵辣椒。将未接种乳酸菌的辣椒为对比试验组。按照GB 5009.33-2010：《食品中亚硝酸盐与硝酸盐的测定》，测定发酵辣椒的亚硝酸盐含量（图4）。按照GB/T 5009.208-2008：《食品中生物胺含量的测定》，使用HPLC法对接种戊糖乳杆菌CICC 6294的发酵辣椒和自然发酵辣椒中生物胺的种类和含量进行测定（图5）。按照GB/T 12456-2008：《食品中总酸的测定》中酸碱滴定法测定发酵辣椒的总酸含量（表1）。
表1 发酵辣椒测定指标
| 测定指标 | 自然发酵 | 戊糖乳杆菌CICC6294 |
| 亚硝酸盐（mg/kg） | 2.63±0.11 | 0.81±0.09 |
| 生物胺（mg/kg） | 19.36±0.24 | 3.19±0.12 |
| 总酸含量（%） | 2.09±0.08 | 2.76±0.05 |
戊糖乳杆菌CICC 6294用于新疆特色辣椒发酵，所得到的发酵辣椒亚硝酸盐的含量0.81 mg/kg, 低于对照组亚硝酸盐含量（2.63 mg/kg），远远低于国家标准20 mg/kg。接种菌株CICC 6294的发酵辣椒中生物胺主要为腐胺，其含量为3.19 mg/kg，大大低于对照组腐胺含量。因此接种戊糖乳杆菌CICC 6294，能够显著降低了发酵蔬菜中残留亚的硝酸盐和生物胺含量，解决了亚硝酸盐和生物胺带来的食品安全问题。此外，接种戊糖乳杆菌CICC6294的发酵辣椒总酸含量2.76%，高于自然发酵辣椒中总酸含量，在生产中能够缩短辣椒发酵的周期。
Claims (4)
1.一种戊糖乳杆菌 CICC 6294，其特征在于：该菌株保藏于中国普通微生物菌种保藏中心，保藏号为CGMCC 14016。
2.根据权利要求1所述的戊糖乳杆菌CICC 6294，其特征在于：该菌株菌体呈短杆状，0.4-0.5 μm × 0.6-1.5 μm，属于革兰氏阳性菌；在改良MRS 液体培养基中培养24 h，亚硝酸盐降解率能够达到98.8%，培养30 h，亚硝酸盐降解率达到100%；该菌株氨基酸脱羧酶为阴性，并且能够抑制大肠埃希氏菌、金黄色葡萄球菌、单增李斯特菌和沙门氏菌致病微生物。
3.根据权利要求1所述的戊糖乳杆菌CICC 6294，其特征在于：菌株CICC 6294可应用于果蔬发酵，尤其是新疆特色辣椒发酵。
4.根据权利要求3所述的用途，其特征在于：接种戊糖乳杆菌CICC 6294发酵的新疆特色辣椒，亚硝酸盐含量为0.81 mg/kg，生物胺的含量为3.19 mg/kg，显著低于国家食品安全标准，极大的提高发酵辣椒的食用安全性。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710328555.2A CN108865919B (zh) | 2017-05-11 | 2017-05-11 | 一种戊糖乳杆菌cicc6294及其在辣椒发酵中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710328555.2A CN108865919B (zh) | 2017-05-11 | 2017-05-11 | 一种戊糖乳杆菌cicc6294及其在辣椒发酵中的应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108865919A true CN108865919A (zh) | 2018-11-23 |
| CN108865919B CN108865919B (zh) | 2021-09-07 |
Family
ID=64319341
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710328555.2A Active CN108865919B (zh) | 2017-05-11 | 2017-05-11 | 一种戊糖乳杆菌cicc6294及其在辣椒发酵中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108865919B (zh) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112029673A (zh) * | 2020-01-14 | 2020-12-04 | 新疆中亚食品研发中心(有限公司) | 一种发酵乳杆菌cicc 6278及其在新疆特色辣椒发酵中的应用 |
| CN116855424A (zh) * | 2023-08-23 | 2023-10-10 | 四川大学 | 一株抑制解脂耶氏酵母菌的戊糖乳植物杆菌及其应用 |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20030064462A (ko) * | 2002-01-28 | 2003-08-02 | (주)바이오토피아 | 유해세균의 성장을 억제하는 신규 미생물 및 이를유효성분으로 하는 미생물 첨가제 |
| CN1566322A (zh) * | 2003-06-26 | 2005-01-19 | 河南双汇投资发展股份有限公司 | 一种戊糖乳杆菌菌株和以该菌株制成的发酵剂及该发酵剂在肉食品中的应用 |
| WO2007003917A1 (en) * | 2005-07-01 | 2007-01-11 | Matforsk As | Probiotic lactobacillus plantarum or pentosus starter strains |
| CN102286392A (zh) * | 2011-03-01 | 2011-12-21 | 安徽农业大学 | 一种戊糖乳杆菌和该乳杆菌的发酵产物以及发酵产物的用途 |
| CN103320337A (zh) * | 2012-10-23 | 2013-09-25 | 云南宏斌绿色食品有限公司 | 戊糖乳杆菌 |
| CN105969680A (zh) * | 2016-03-23 | 2016-09-28 | 贵州大学 | 一株降胆固醇、降亚硝酸盐的戊糖乳杆菌及其筛选方法 |
-
2017
- 2017-05-11 CN CN201710328555.2A patent/CN108865919B/zh active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20030064462A (ko) * | 2002-01-28 | 2003-08-02 | (주)바이오토피아 | 유해세균의 성장을 억제하는 신규 미생물 및 이를유효성분으로 하는 미생물 첨가제 |
| CN1566322A (zh) * | 2003-06-26 | 2005-01-19 | 河南双汇投资发展股份有限公司 | 一种戊糖乳杆菌菌株和以该菌株制成的发酵剂及该发酵剂在肉食品中的应用 |
| WO2007003917A1 (en) * | 2005-07-01 | 2007-01-11 | Matforsk As | Probiotic lactobacillus plantarum or pentosus starter strains |
| CN102286392A (zh) * | 2011-03-01 | 2011-12-21 | 安徽农业大学 | 一种戊糖乳杆菌和该乳杆菌的发酵产物以及发酵产物的用途 |
| CN103320337A (zh) * | 2012-10-23 | 2013-09-25 | 云南宏斌绿色食品有限公司 | 戊糖乳杆菌 |
| CN105969680A (zh) * | 2016-03-23 | 2016-09-28 | 贵州大学 | 一株降胆固醇、降亚硝酸盐的戊糖乳杆菌及其筛选方法 |
Non-Patent Citations (4)
| Title |
|---|
| FANGDA SUN等: "N-nitrosoamine inhibition and quality preservation of Harbin dry sausages by inoculated with Lactobacillus pentosus, Lactobacillus curvatus and Lactobacillus sake", 《FOOD CONTROL》 * |
| PING-MEI YAN等: "Effect of inoculating lactic acid bacteria starter cultures on the nitrite concentration of fermenting Chinese paocai", 《FOOD CONTROL》 * |
| 宋小娟: "适用于肉制品发酵的降胆固醇乳酸菌的筛选及性能评价", 《中国优秀硕士学位论文全文数据库 工程科技I辑》 * |
| 翟磊等: "菌株CICC 6294生物学特性及其在辣椒发酵中的应用", 《食品发酵与工业》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112029673A (zh) * | 2020-01-14 | 2020-12-04 | 新疆中亚食品研发中心(有限公司) | 一种发酵乳杆菌cicc 6278及其在新疆特色辣椒发酵中的应用 |
| CN112029673B (zh) * | 2020-01-14 | 2023-08-08 | 新疆新康农业发展有限公司 | 一种发酵乳杆菌cicc 6278及其在新疆特色辣椒发酵中的应用 |
| CN116855424A (zh) * | 2023-08-23 | 2023-10-10 | 四川大学 | 一株抑制解脂耶氏酵母菌的戊糖乳植物杆菌及其应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108865919B (zh) | 2021-09-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104531578B (zh) | 具有高效降解亚硝酸盐和强产酸功能的植物乳杆菌菌株及其用途 | |
| CN106754472B (zh) | 一株果蔬发酵用植物乳杆菌grx15及其应用 | |
| CN104593290B (zh) | 一种具有抗氧化功能的泡菜复合发酵剂配方及其应用 | |
| CN103013861B (zh) | 枯草芽孢杆菌hjda32菌株及其所产细菌素的制备方法 | |
| CN103509734B (zh) | 一种果酒生物降酸新菌株、制备方法及其应用 | |
| CN104357353A (zh) | 一株植物乳杆菌在发酵果蔬汁制作中的应用 | |
| CN104164459A (zh) | 利用发酵提高糙米γ—氨基丁酸的方法 | |
| CN101967454A (zh) | 一种乳酸菌高密度培养方法 | |
| CN107172883A (zh) | 瓜氨酸的制备方法 | |
| CN103571782A (zh) | 一株融合乳杆菌及其应用 | |
| CN105132321A (zh) | 一种屎肠球菌及其高密度固态发酵的培养基及方法 | |
| CN113980853B (zh) | 一株产乳酸的格氏乳球菌wbt0008及其应用 | |
| CN106635916B (zh) | 东方醋酸菌yzd-09及其应用 | |
| CN107034161B (zh) | 一种产马乳酒乳杆菌及其在辣椒发酵中的应用 | |
| CN1670184A (zh) | 肽类天然微生物防腐剂产生菌及应用和防腐剂的制备方法 | |
| CN104877940B (zh) | 一株嗜热链球菌 | |
| CN108865919A (zh) | 一种戊糖乳杆菌cicc6294及其在辣椒发酵中的应用 | |
| CN112029673B (zh) | 一种发酵乳杆菌cicc 6278及其在新疆特色辣椒发酵中的应用 | |
| CN114231458B (zh) | 一种提高瓜果类糖酸比的复合微生物菌剂及其制备方法和应用 | |
| CN110257306A (zh) | 一株植物乳杆菌及其应用 | |
| CN116024133A (zh) | 一株耐高浓度苹果酸的植物乳杆菌及其应用 | |
| CN106591176B (zh) | 一种戊糖乳杆菌及其应用 | |
| CN108546654B (zh) | 一种降酸发酵乳杆菌及其应用 | |
| KR101744529B1 (ko) | 바실러스 서브틸리스 1-d-5, 이를 이용한 비피도박테리아 증식방법 및 효소식품 제조방법 | |
| CN107189959A (zh) | 瑞士乳杆菌grx09及其应用 |
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 | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20220118 Address after: 830032 factory 2, comprehensive free trade zone, No. 9, tunping North Road, Xinjiang Urumqi Economic and Technological Development Zone (Toutunhe District), Urumqi City, Xinjiang Uygur Autonomous Region Patentee after: XINJIANG CENTRAL ASIA FOOD RESEARCH AND DEVELOPMENT CENTER (CO.,LTD.) Patentee after: CHINA NATIONAL RESEARCH INSTITUTE OF FOOD & FERMENTATION INDUSTRIES Co.,Ltd. Address before: 830026 Satellite Road 533, Urumqi economic and Technological Development Zone, the Xinjiang Uygur Autonomous Region Patentee before: XINJIANG CENTRAL ASIA FOOD RESEARCH AND DEVELOPMENT CENTER (CO.,LTD.) |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20220630 Address after: No.2 factory building, comprehensive bonded zone, No.9 tunping North Road, Urumqi Economic and Technological Development Zone (Toutunhe District), 830000 Xinjiang Uygur Autonomous Region Patentee after: Xinjiang Xinkang Agricultural Development Co.,Ltd. Address before: 830032 factory 2, comprehensive free trade zone, No. 9, tunping North Road, Xinjiang Urumqi Economic and Technological Development Zone (Toutunhe District), Urumqi City, Xinjiang Uygur Autonomous Region Patentee before: XINJIANG CENTRAL ASIA FOOD RESEARCH AND DEVELOPMENT CENTER (CO.,LTD.) Patentee before: CHINA NATIONAL RESEARCH INSTITUTE OF FOOD & FERMENTATION INDUSTRIES Co.,Ltd. |
|
| TR01 | Transfer of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A Lactobacillus pentosus CICC6294 and its application in chili fermentation Effective date of registration: 20231024 Granted publication date: 20210907 Pledgee: Bank of China Limited Xinjiang Uygur Autonomous Region Branch Pledgor: Xinjiang Xinkang Agricultural Development Co.,Ltd. Registration number: Y2023650000010 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |