产氨基甲酸乙酯降解酶酵母菌的筛选鉴定及发酵条件优化
|
摘要
采用选择性培养基,从36株酵母菌中分离纯化得到4株(Y4、Y9、Y13、N6)产氨基甲酸乙酯降解酶酵母菌,通过ITS序列比对,均属于Pichia kudriavzevii。通过单因素试验和正交试验,对Y13菌株产EC水解酶的发酵条件进行优化。结果表明:最优条件是以10g/L麦芽糖为碳源;以30g/L酵母浸粉为氮源;接种量为6%;培养基中EC的底物浓度为5.00%,装液量为20%,初始pH为6,培养温度为28℃,培养时间为120h。
Four strains(Y4,Y9,Y13,N6)of yeasts producing EC-degrading enzymes are isolated and purified from 36 strains of yeasts by selective medium.The ITS sequence alignments belong to Pichia kudriavzevii.The fermentation conditions of EC hydrolase produced by Y13 strain are optimized by single factor test and orthogonal test.The results show that the optimum conditions are 10 g/L maltose as the carbon source;30 g/L yeast extract as the nitrogen source;the inoculum size is 6%;the substrate concentration of EC in the medium is 5.00%,the liquid volume is 20%,the initial pH is 6,the culture temperature is 28℃,and the culture time is 120 h.
Four strains(Y4,Y9,Y13,N6)of yeasts producing EC-degrading enzymes are isolated and purified from 36 strains of yeasts by selective medium.The ITS sequence alignments belong to Pichia kudriavzevii.The fermentation conditions of EC hydrolase produced by Y13 strain are optimized by single factor test and orthogonal test.The results show that the optimum conditions are 10 g/L maltose as the carbon source;30 g/L yeast extract as the nitrogen source;the inoculum size is 6%;the substrate concentration of EC in the medium is 5.00%,the liquid volume is 20%,the initial pH is 6,the culture temperature is 28℃,and the culture time is 120 h.
引文
[1]Dennis M J,Howarth N,Key P E,et al.Investigation of ethyl carbamate levels in some fermented foods and alcoholic beverages[J].Food Additives and Contaminants,1989,6(3):383-389.
[2]Kobashi K.Ethyl carbamate in alcoholic beverages[J].Eiseikagaku,1989,35(2):110-124.
[3]Ough C S.Ethyl carbamate in fermented beverages and foods.I.Naturally occurring ethylcarbamate[J].Journal of Agricultural and Food Chemistry,1976,24(2):323-328.
[4]Schmeltz I,Chiong K G,Hoffmann D.Formation and determination of ethyl carbamate in tobacco and tobacco smoke[J].Journal of Analytical Toxicology,1978,2(6):265-268.
[5]陆健,曹任.葡萄酒中氨基甲酸乙酯的研究[J].食品与发酵工业,1996(3):79-80.
[6]李伍林,汪新亮,周平春.氨基甲酸乙酯的合成[J].咸宁学院学报,2005(6):99-100.
[7]王德良,王晓娟,傅力,等.复合诱变选育低产尿素黄酒酵母[J].酿酒科技,2009(8):17-21.
[8]李京京,方芳,张继冉,等.氨基甲酸乙酯水解酶的分离纯化及酶学性质[J].食品与生物技术学报,2014,33(12):1239-1245.
[9]Guo X,Li Y,Guo J,et al.Reduced production of ethyl carbamate for wine fermentation by deleting CAR1 in Saccharomyces cerevisiae[J].J Ind Microbiol Biotechnol,2016,43:671-679.
[10]张俊杰,杨旭,张文叶,等.河南太行山区野生山葡萄表皮酵母菌的分离与鉴定[J].酿酒科技,2016(5):57-60,64.
[11]Matsushita K,Noue T,Adachi O,et al.Acetobacter aceti possesses a proton motive force-dependent efflux system for acetic acid[J].J Bacteriol,2005,187(13):4346-4352.
[12]Beijerinck M.Ueber die arten der essigbakterien[J].Centralblatt für Bakteriologie,Parasitenkunde and Infektions Krankheiten,2006,2(4):209-216.
[13]沈萍,陈向东.微生物学实验[M].北京:高等教育出版社,2007:81-84.
[14]Hossein K,Ladan K,Nilufar J,et al.Utilization of size polymorphism in ITS1and ITS2regions for identification of pathogenic yeast species[J].Journal of Medical Microbiology,2017,66:126-133.
[2]Kobashi K.Ethyl carbamate in alcoholic beverages[J].Eiseikagaku,1989,35(2):110-124.
[3]Ough C S.Ethyl carbamate in fermented beverages and foods.I.Naturally occurring ethylcarbamate[J].Journal of Agricultural and Food Chemistry,1976,24(2):323-328.
[4]Schmeltz I,Chiong K G,Hoffmann D.Formation and determination of ethyl carbamate in tobacco and tobacco smoke[J].Journal of Analytical Toxicology,1978,2(6):265-268.
[5]陆健,曹任.葡萄酒中氨基甲酸乙酯的研究[J].食品与发酵工业,1996(3):79-80.
[6]李伍林,汪新亮,周平春.氨基甲酸乙酯的合成[J].咸宁学院学报,2005(6):99-100.
[7]王德良,王晓娟,傅力,等.复合诱变选育低产尿素黄酒酵母[J].酿酒科技,2009(8):17-21.
[8]李京京,方芳,张继冉,等.氨基甲酸乙酯水解酶的分离纯化及酶学性质[J].食品与生物技术学报,2014,33(12):1239-1245.
[9]Guo X,Li Y,Guo J,et al.Reduced production of ethyl carbamate for wine fermentation by deleting CAR1 in Saccharomyces cerevisiae[J].J Ind Microbiol Biotechnol,2016,43:671-679.
[10]张俊杰,杨旭,张文叶,等.河南太行山区野生山葡萄表皮酵母菌的分离与鉴定[J].酿酒科技,2016(5):57-60,64.
[11]Matsushita K,Noue T,Adachi O,et al.Acetobacter aceti possesses a proton motive force-dependent efflux system for acetic acid[J].J Bacteriol,2005,187(13):4346-4352.
[12]Beijerinck M.Ueber die arten der essigbakterien[J].Centralblatt für Bakteriologie,Parasitenkunde and Infektions Krankheiten,2006,2(4):209-216.
[13]沈萍,陈向东.微生物学实验[M].北京:高等教育出版社,2007:81-84.
[14]Hossein K,Ladan K,Nilufar J,et al.Utilization of size polymorphism in ITS1and ITS2regions for identification of pathogenic yeast species[J].Journal of Medical Microbiology,2017,66:126-133.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |