山西老陈醋中芽孢杆菌的RAPD分型及其与性状之间的关系
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摘要
从山西老陈醋醋酸发酵过程的醋醅中分离芽孢杆菌(Bacillus),采用分子生物学技术对其进行鉴定,并研究菌株随机扩增多态性脱氧核糖核酸(RAPD)分型和产酶、乙偶姻、多酚、总酯之间的关系,从中筛选一株优良菌株。结果表明,共分离出30株芽孢杆菌,其中11株地衣芽孢杆菌(Bacillus licheniformis)、8株解淀粉芽孢杆菌(Bacillus amyloliquefaciens)、6株枯草芽孢杆菌(Bacillus subtilis)、2株甲基营养型芽孢杆菌(Bacillus methylotrophicus)和短小芽孢杆菌(Bacillus pumilus)、1株沙福芽孢杆菌(Bacillus safensis)。芽孢杆菌种内具有基因多样性,同种同型别的芽孢杆菌性状具有相似性;但同种不同型别的芽孢杆菌特性具有差异性。最终筛选出1株产糖化酶(89.68 U/m L)、蛋白酶(34.53 U/m L)、乙偶姻(1.05 mg/m L)、多酚(401.36μg/m L)、总酯(9.62 g/100 m L)能力均较强的解淀粉芽孢杆菌38。
Bacillus was isolated from Cupei in the acetic acid fermentation process of Shanxi aged vinegar, and identified by molecular biotechnology.A good strain was selected by studying the relationship between random amplified polymorphic deoxyribonucleic acid(RAPD) typing and enzyme,acetoin, polyphenols, and total esters production. The results showed that 30 strains of Bacillus were isolated, including 11 strains of Bacillus licheniformis, 8 strains of Bacillus amyloliquefaciens, 6 strains of Bacillus subtilis, 2 strains of Bacillus methylotrophicus and Bacillus pumilus, and 1 strain of Bacillus safensis. The Bacillus species had genetic diversity, and the traits of same species and type of Bacillus had similarity; the Bacillus of same species but different types had different characteristics. Finally, B. amyloliquefaciens 38 with strong ability of producing saccharification enzyme(89.68 U/ml), protease(34.53 U/ml), acetoin(1.05 mg/ml), polyphenols(401.36 μg/ml) and total ester(9.62 g/100 ml) was screened out.
Bacillus was isolated from Cupei in the acetic acid fermentation process of Shanxi aged vinegar, and identified by molecular biotechnology.A good strain was selected by studying the relationship between random amplified polymorphic deoxyribonucleic acid(RAPD) typing and enzyme,acetoin, polyphenols, and total esters production. The results showed that 30 strains of Bacillus were isolated, including 11 strains of Bacillus licheniformis, 8 strains of Bacillus amyloliquefaciens, 6 strains of Bacillus subtilis, 2 strains of Bacillus methylotrophicus and Bacillus pumilus, and 1 strain of Bacillus safensis. The Bacillus species had genetic diversity, and the traits of same species and type of Bacillus had similarity; the Bacillus of same species but different types had different characteristics. Finally, B. amyloliquefaciens 38 with strong ability of producing saccharification enzyme(89.68 U/ml), protease(34.53 U/ml), acetoin(1.05 mg/ml), polyphenols(401.36 μg/ml) and total ester(9.62 g/100 ml) was screened out.
引文
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[23]黄家驷,刘盾,陈玄阳,等.浓香型高温酒曲中蛋白酶产生菌的筛选与鉴定[J].安徽农学通报,2017(21):35-37.
[24]董丹,车振明,关统伟.白酒酒糟中产纤维素酶菌株的筛选及其酶活力特性检测[J].中国酿造,2015,34(8):44-48.
[25]王霜,缪礼鸿,张明春,等.浓酱兼香型酒醅中产酱香芽孢杆菌的筛选及发酵风味成分分析[J].中国酿造,2017,36(10):61-65.
[2]王进龙.山西老陈醋大曲中芽孢杆菌的分离鉴定及特性研究[D].晋中:山西农业大学,2016.
[3]VIDAL A,MARIN S,SANCHIS V,et al.Hydrolysers of modified mycotoxins in maize:α-amylase and cellulase induce an underestimation of the total aflatoxin content[J].Food Chem,2018,248(6):86-92.
[4]史改玲.山西老陈醋源优良芽孢杆菌菌株的鉴定及筛选[J].中国酿造,2018,37(5):22-27.
[5]AKASAKA,NAOKI,SAKODA,et al.An efficient method using gluconacetobacter europaeus to reduce an unfavorable flavor compound,acetoin,in rice vinegar production[J].Appl Environ Microbiol,2013,79(23):7334-7342.
[6]LU L,WEI X,YU W,et al.Enhancement of acetoin production from Bacillus licheniformis by 2,3-butanediol conversion strategy:Metabolic engineering and fermentation control[J].Process Biochem,2017,57(10):35-42.
[7]高文睿,贠建民.凉州熏醋中高产乙偶姻菌株的分离筛选及发酵特性初探[J].农业科技与信息,2015(16):26-29.
[8]张显.高产乙偶姻枯草芽孢杆菌的代谢工程改造[D].无锡:江南大学,2013.
[9]郇阿梅,刘军,王洋,等.一株分离至传统麸醋醋醅芽孢杆菌发酵特性研究[J].中国调味品,2014,39(8):23-28.
[10]ENAN M R.Application of random amplified polymorphic DNA(RAPD)to detect the genotoxic effect of heavy metals[J].Biotechnol Appl Bioc,2011,43(3):147-154.
[11]李艳莉.酸奶及发酵剂中分离乳酸菌药物敏感性研究[D].广州:华南理工大学,2015.
[12]TAKUYA Y,TATSUJI S,TAKENORI N,et al.Lactic acid bacteria from kefir increase cytotoxicity of natural killer cells to tumor cells[J].Foods,2018,7(4):48-56.
[13]KHOWAL S,SIDDIQUI M Z,ALI S,et al.A report on extensive lateral genetic reciprocation between arsenic resistant Bacillus subtilis and Bacillus pumilus strains analyzed using RAPD-PCR[J].Mol Phylogenet Evol,2017,107:443-454.
[14]RAMALINGAM V,RAJARAM R,ANANTH S,et al.Development of thermostable amylase enzyme from Bacillus cereus for potential antibiofilm activity[J].Bioorg Chem,2018,77:494-506.
[15]MIKHAIL M.VOROB'EV,BUTRE C I,SFORZA S,et al.Demasking kinetics of peptide bond cleavage for whey protein isolate hydrolysed by Bacillus licheniformis protease[J].J Mol Catal B-Enzym,2017,133(1):26-31.
[16]ALVAREZJUBETE L,WIJNGAARD H,ARENDT E K,et al.Polyphenol composition and in vitro antioxidant activity of amaranth,quinoa buckwheat and wheat as affected by sprouting and baking[J].Food Chem,2010,119(2):770-778.
[17]SENTER S D,CHAPMAN G W,WRJR F,et al.Sugar and nonvolatile acid composition of persimmons during maturation[J].J Food Sci,2010,56(4):989-991.
[18]CIANI M,COMITINI F,MANNAZZU I,et al.Controlled mixed culture fermentation:a new perspective on the use of non-Saccharomyces yeasts in winemaking[J].Fems Yeast Res,2010,10(2):123-133.
[19]CIHAN A C,YILDIZ E D,SAHIN E,et al.Introduction of novel thermostableα-amylases from genus anoxybacillus and proposing to group the Bacillaceae relatedα-amylases under five individual GH13 subfamilies[J].World J Microb Biot,2018,34(7):95.
[20]乔羽,于迪,邢晓莹.山西老陈醋发酵阶段高产淀粉酶芽孢杆菌的筛选与鉴定[J].中国酿造,2018,37(1):30-34.
[21]徐颖.高产α-淀粉酶生产菌的筛选鉴定及其酶学性质研究[D].成都:四川大学,2007.
[22]贾莉,郝林,金越.产酸性蛋白酶菌株在老陈醋澄清上的应用[J].食品工程,2003(2):22-23.
[23]黄家驷,刘盾,陈玄阳,等.浓香型高温酒曲中蛋白酶产生菌的筛选与鉴定[J].安徽农学通报,2017(21):35-37.
[24]董丹,车振明,关统伟.白酒酒糟中产纤维素酶菌株的筛选及其酶活力特性检测[J].中国酿造,2015,34(8):44-48.
[25]王霜,缪礼鸿,张明春,等.浓酱兼香型酒醅中产酱香芽孢杆菌的筛选及发酵风味成分分析[J].中国酿造,2017,36(10):61-65.
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