1株能利用高粱汁产L-乳酸的菌株鉴定及培养基优化
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摘要
以产L-乳酸的菌株A2为对象,采用16S rRNA基因测序法结合菌株的表型特征进行鉴定,以甜高粱汁为主要培养基质,采用响应面设计软件对该菌种的发酵培养基进行优化。结果表明,A2菌株为Lactobacillus plantarum S4,优化得到的甜高粱汁培养基配比为甜高粱汁327. 83 g/L,蛋白胨1. 67 g/L,磷酸氢二钾4. 7 g/L,硫酸锰0. 17 g/L,在此培养基配比下,Lactobacillus plantarum S4厌氧发酵68 h后,L-乳酸产量达(61. 20±1. 36) g/L,L-乳酸/葡萄糖转化率(90. 74±2. 28)%,L-乳酸/蔗糖转化率(47. 20±1. 81)%。
In this study a L-lactic acid producing strain A2 was used as a target to carry out phenotypic characterization adopting 16 S rRNA gene sequencing,and carried out the fermentation medium optimization taking sweet sorghum juice as main culturing stroma,and using Response Surface Designing Software. The results showed that the strain A2 was Lactobacillus plantarum S4. The optimized media proportion of different elements contained: sweet sorghum juice327. 83 g/L,peptone 1. 67 g/L,K2 HPO44. 7 g/L,MnSO4·4 H2 O 0. 17 g/L. Based on this proportion of media,68 h after anaerobic fermentation,the yield of L-lactic acid was as high as( 61. 20 ± 1. 36) g/L,the L-lactic acid/glucose conversion rate was( 90. 74 ± 2. 28) %,L-lactic acid/sucrose conversion rate was( 47. 20 ± 1. 81) %.
In this study a L-lactic acid producing strain A2 was used as a target to carry out phenotypic characterization adopting 16 S rRNA gene sequencing,and carried out the fermentation medium optimization taking sweet sorghum juice as main culturing stroma,and using Response Surface Designing Software. The results showed that the strain A2 was Lactobacillus plantarum S4. The optimized media proportion of different elements contained: sweet sorghum juice327. 83 g/L,peptone 1. 67 g/L,K2 HPO44. 7 g/L,MnSO4·4 H2 O 0. 17 g/L. Based on this proportion of media,68 h after anaerobic fermentation,the yield of L-lactic acid was as high as( 61. 20 ± 1. 36) g/L,the L-lactic acid/glucose conversion rate was( 90. 74 ± 2. 28) %,L-lactic acid/sucrose conversion rate was( 47. 20 ± 1. 81) %.
引文
[1] Glaser R,Venus J. Model-based characterisation of growth performance and L-lactic acid production with high optical purity by thermophilic Bacillus coagulans in a lignin-supplemented mixed substrate medium[J]. New Biotech,2017,37:180-193.
[2] Castillo Martinez FA,Balciunas EM,Salgado JM,et al. Lactic acid properties,applications and production:a review[J].Trends Food Sci Technol.,2013,30:70-83.
[3] Coutu DL,Yousefi AM,Galipeau J. Three-dimensional porous scaffolds at the crossroads of tissue engineering and cell-based gene therapy[J]. J Cell Biochem.,2009,15:537-546.
[4] Gupta B,Revagade N,Hilborn J. Poly(lactic acid)fiber:an overview[J]. Prog Polym Sci.,2007,34:455-482.
[5]丁涓,魏敏,张莉.玉米浆发酵生产L-乳酸的工艺优化[J].食品科学,2011,32(1):127-130.
[6]于波,曾艳,姜旭,等.聚合级L-乳酸的非粮生物质发酵研究进展[J].生物工程学报,2013,29(4):411-421.
[7]姚纲,张健鹏,胡红焱,等.用API50 CH系统鉴定9株产高光学纯度D-乳酸野生菌株[J].中国实用医药,2014,9(8):263-264.
[8]袁世超.优良乳酸菌的选育及在保育猪生产中的应用研究[D].郑州:郑州大学,2015.
[9]段宇珩,谈重芳,王雁萍,等.乳酸菌鉴定方法在食品工业中的应用及研究进展[J].食品工业科技,2007,28(2):242-244.
[10]尹胜利,杜鉴,徐晨.乳酸菌的研究现状及其应用[J].食品科技,2012,37(9):25-29.
[11]凌代文.乳酸菌的分类鉴定及实验方法[M].北京:中国轻工业出版社,1999.
[12]周广麒,翟景先.甜高粱秸秆汁的成分及其乙醇发酵的研究[J].生产与科研经验,2010,36(1):85-87.
[13] Evaggcli B,Dimitris PK,Bernard M,et al. Structure and composition of sweet sorghum stalk components[J]. Industrial Crops and Products,1997,6(4):297-302.
[14] Lane DJ. 16S/23S rRNA sequencing. In:Stackebrandt,E.,Goodfellow,M.(Eds.),Nucleic Acid Techniques in Bacterial Systematics[N]. John Wiley&Sons,Chichester,pp.,1991:115-175.
[15] Tamura K,Peterson D,Peterson N,et al. MEGA5:molecular evolutionary genetics analysis using maximum likelihood,evolutionary distance, and maximum parsimony methods[J].Mol. Biol. Evol.,2011,28:2731-2739.
[16]洪庆华,迟杰,齐云,等.影响革兰染色结果的原因分析[J].实验技术与管理,2011,5:41-43.
[17]马蓓,刘伟,谢蓉蓉,等.乳酸菌扫描电镜制样方法及观察条件探究[J].内蒙古科技与经济,2012,252(2):116-119.
[18] Ouyang J,Ma R,Zheng ZJ,et al. Open fermentative production of L-lactic acid by Bacillus sp. strain NL01 using lignocellulosic hydrolyzates as low-cost raw material[J]. Bioresource Technol.,2013,135:475-480.
[19] Gao T,Wong Y,Ng C,et al. L-Lactic acid production by Bacillus subtilis MUR1[J]. Bioresource. Technol.,2012,121:105-110.
[20] Qin JY,Wang XW,Zheng ZJ,et al. Production of L-lactic acid by a thermophilic Bacillus mutant using sodium hydroxide as neutralizing agent[J]. Bioresource Technol.,2010,101:7570-7576.
[21] Zhan X,Wang D,Tuinstra MR,et al. Ethanol and lactic acid production as affected by sorghum genotype and location[J].Ind Crop Prod,2003,18:245-255.
[22] Sule B,Murat E,Dursun O. Effect of different carbon sources on L(+)-lactic acid production by Rhizopus oryzae[J]. Biochem Eng J,2004,21:33-37.
[23] Gao T,Wong Y,Ng C,et al. L-lactic acid production by Bacillus subtilis MUR1[J]. Bioresource Technol,2012,121:105-110.
[2] Castillo Martinez FA,Balciunas EM,Salgado JM,et al. Lactic acid properties,applications and production:a review[J].Trends Food Sci Technol.,2013,30:70-83.
[3] Coutu DL,Yousefi AM,Galipeau J. Three-dimensional porous scaffolds at the crossroads of tissue engineering and cell-based gene therapy[J]. J Cell Biochem.,2009,15:537-546.
[4] Gupta B,Revagade N,Hilborn J. Poly(lactic acid)fiber:an overview[J]. Prog Polym Sci.,2007,34:455-482.
[5]丁涓,魏敏,张莉.玉米浆发酵生产L-乳酸的工艺优化[J].食品科学,2011,32(1):127-130.
[6]于波,曾艳,姜旭,等.聚合级L-乳酸的非粮生物质发酵研究进展[J].生物工程学报,2013,29(4):411-421.
[7]姚纲,张健鹏,胡红焱,等.用API50 CH系统鉴定9株产高光学纯度D-乳酸野生菌株[J].中国实用医药,2014,9(8):263-264.
[8]袁世超.优良乳酸菌的选育及在保育猪生产中的应用研究[D].郑州:郑州大学,2015.
[9]段宇珩,谈重芳,王雁萍,等.乳酸菌鉴定方法在食品工业中的应用及研究进展[J].食品工业科技,2007,28(2):242-244.
[10]尹胜利,杜鉴,徐晨.乳酸菌的研究现状及其应用[J].食品科技,2012,37(9):25-29.
[11]凌代文.乳酸菌的分类鉴定及实验方法[M].北京:中国轻工业出版社,1999.
[12]周广麒,翟景先.甜高粱秸秆汁的成分及其乙醇发酵的研究[J].生产与科研经验,2010,36(1):85-87.
[13] Evaggcli B,Dimitris PK,Bernard M,et al. Structure and composition of sweet sorghum stalk components[J]. Industrial Crops and Products,1997,6(4):297-302.
[14] Lane DJ. 16S/23S rRNA sequencing. In:Stackebrandt,E.,Goodfellow,M.(Eds.),Nucleic Acid Techniques in Bacterial Systematics[N]. John Wiley&Sons,Chichester,pp.,1991:115-175.
[15] Tamura K,Peterson D,Peterson N,et al. MEGA5:molecular evolutionary genetics analysis using maximum likelihood,evolutionary distance, and maximum parsimony methods[J].Mol. Biol. Evol.,2011,28:2731-2739.
[16]洪庆华,迟杰,齐云,等.影响革兰染色结果的原因分析[J].实验技术与管理,2011,5:41-43.
[17]马蓓,刘伟,谢蓉蓉,等.乳酸菌扫描电镜制样方法及观察条件探究[J].内蒙古科技与经济,2012,252(2):116-119.
[18] Ouyang J,Ma R,Zheng ZJ,et al. Open fermentative production of L-lactic acid by Bacillus sp. strain NL01 using lignocellulosic hydrolyzates as low-cost raw material[J]. Bioresource Technol.,2013,135:475-480.
[19] Gao T,Wong Y,Ng C,et al. L-Lactic acid production by Bacillus subtilis MUR1[J]. Bioresource. Technol.,2012,121:105-110.
[20] Qin JY,Wang XW,Zheng ZJ,et al. Production of L-lactic acid by a thermophilic Bacillus mutant using sodium hydroxide as neutralizing agent[J]. Bioresource Technol.,2010,101:7570-7576.
[21] Zhan X,Wang D,Tuinstra MR,et al. Ethanol and lactic acid production as affected by sorghum genotype and location[J].Ind Crop Prod,2003,18:245-255.
[22] Sule B,Murat E,Dursun O. Effect of different carbon sources on L(+)-lactic acid production by Rhizopus oryzae[J]. Biochem Eng J,2004,21:33-37.
[23] Gao T,Wong Y,Ng C,et al. L-lactic acid production by Bacillus subtilis MUR1[J]. Bioresource Technol,2012,121:105-110.