富硒酵母的分离鉴定及富硒条件的优化
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摘要
富硒酵母生产发酵食品是补充硒元素的有效方法,以实验室保存的酵母菌为材料,通过选择培养基平板稀释法,分离纯化得到四株富硒酵母,经形态学和分子生物学鉴定,初步鉴定菌株Z4、Z5、Z7、Z9均为库德毕赤酵母(Pichia kudriavzevii)。通过对比四株菌的生长曲线及富硒能力,发现Z5菌株生长状况最好,选取接种量、碳源(葡萄糖)和氮源(蛋白胨)添加量作为单一因素,通过单因素试验和正交试验,探究各影响因素对菌株Z5富硒能力的影响。结果表明,Z5菌株的最佳富硒发酵条件为接种量10%,葡萄糖2.4%,蛋白胨1.8%。在此最佳条件下,菌株Z5生物量为9.63 g/L,总硒含量可达到862μg/g。
Fermented food produced by selenium-enriched yeast is an effective method for supplementing selenium. Four selenium-enriched yeasts were isolated from the yeast stored in the laboratory and purified by selective medium dilution method, after morphology and molecular biology identification, the results showed that strain Z4, Z5, Z7 and Z9 were identified as Pichia kudriavzevii. The strain Z5 with good growth condition was obtained by comparing the growth curves and selenium enrichment capacities of the four strains. The inoculum, carbon source(glucose) and nitrogen source(peptone) addition were selected as single factors, and the effect of each influencing factors on selenium-enriching ability of the strain Z5 was investigated by single factor and orthogonal experiments. The results showed that the optimal selenium-enriched conditions of strain Z5 were inoculum10%, glucose 2.4% and peptone 1.8%. Under the optimal conditions, the biomass and total selenium content of strain Z5 was 9.63 g/L and 862 μg/g,respectively.
Fermented food produced by selenium-enriched yeast is an effective method for supplementing selenium. Four selenium-enriched yeasts were isolated from the yeast stored in the laboratory and purified by selective medium dilution method, after morphology and molecular biology identification, the results showed that strain Z4, Z5, Z7 and Z9 were identified as Pichia kudriavzevii. The strain Z5 with good growth condition was obtained by comparing the growth curves and selenium enrichment capacities of the four strains. The inoculum, carbon source(glucose) and nitrogen source(peptone) addition were selected as single factors, and the effect of each influencing factors on selenium-enriching ability of the strain Z5 was investigated by single factor and orthogonal experiments. The results showed that the optimal selenium-enriched conditions of strain Z5 were inoculum10%, glucose 2.4% and peptone 1.8%. Under the optimal conditions, the biomass and total selenium content of strain Z5 was 9.63 g/L and 862 μg/g,respectively.
引文
[1]杨善岩,李海龙,狄志鸿.硒元素生理功能及微生物富硒发酵研究现状[J].食品工业,2013,34(6):167-170.
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[14]DE JONG J V,MCLEOD J C,STEEL M,et al.Neighbourhoods of phylogenetic trees:exact and asymptotic counts[J].SIAM J Discrete Math,2016,30(4):2265-2287.
[15]贺莹,赵建英,母焱楠.响应面法优化富硒酵母的营养条件优化[J].食品工业,2017,38(12):30-34.
[16]孙平平.富硒酵母的选育、发酵条件优化及其应用研究[D].芜湖:安徽工程大学,2011.
[2]YANG L S,WANG W Y,HOU S F,et al.Effects of selenium supplementation on arsenism:an intervention trial in Inner Mongolia[J].Environ Geochem Health,2004,24(4):359-374.
[3]SCHARTZ K,FOLTZ C M.Selenium as an intergral part of factor 3against dietary necrotic liver degeneration[J].Am Chem Soc,2007,79(6):3292-3293.
[4]CASALONE E,DI ILIO C,FEDERICI G,et al.Glutathione and glutathione metabolizing enzymes in yeasts[J].Anton Leeuw,2004,54(4):367-375.
[5]史丽英.人体必需微量元素---硒[J].微量元素与健康研究,2005,22(4):61-63.
[6]迟淑艳,张伟力.微量元素硒与碘的生物学功能研究进展[J].饲料博览,2003,15(3):5-7.
[7]周琼,赵松.富硒酵母中有机硒含量测量方法的探究[J].广州化工,2017,45(21):121-123.
[8]肖方正,刘曲滨.富硒酵母的研究开发与应用[J].广东微量元素科学,2001,8(8):7-9.
[9]刘杰,徐林,吴根福.富硒酵母的研究进展[J].饲料工业,2009,30(22):44-47.
[10]苗君.酿酒酵母在极端条件下耐受性的研究[D].大连:大连工业大学,2009.
[11]巴尼特.酵母菌的特征与鉴定手册[M].青岛:青岛海洋大学出版社,1991:3-10.
[12]BAKER S,NICKLIA J,KHAN N,et al.Microbiology[M].Beijing:Science Press,2013.
[13]ANDORRA I,ESTEVE-ZARZOSO B,GUILLAMON J M.Determination of viable wine yeast using DNA binding dyes and quantitative PCR[J].Int J Food Microbiol,2010,144(2):257-262.
[14]DE JONG J V,MCLEOD J C,STEEL M,et al.Neighbourhoods of phylogenetic trees:exact and asymptotic counts[J].SIAM J Discrete Math,2016,30(4):2265-2287.
[15]贺莹,赵建英,母焱楠.响应面法优化富硒酵母的营养条件优化[J].食品工业,2017,38(12):30-34.
[16]孙平平.富硒酵母的选育、发酵条件优化及其应用研究[D].芜湖:安徽工程大学,2011.