细菌纤维素产生菌选育与突变株发酵特性的研究
|
摘要
为了获得细菌纤维素高产菌株,采用紫外线和硫酸二乙酯对实验室保存的木葡糖酸醋杆菌BC13进行复合诱变。通过诱变、筛选和连续传代实验,获得1株产量达15.6g/L的高产突变株Y19-11,该菌株遗传稳定性良好,比原出发菌株产量提高44.4%。同时,通过比较不同温度、不同初始pH下的菌体量和细菌纤维素产量,确定突变株Y19-11的最适温度和初始pH分别为30℃和5.5。
In order to obtain high bacterial cellulose(BC)-producing strain,composite mutagenesis of ultraviolet and diethyl sulfate was used to Gluconacetobacter xylinus BC13 which preserved in laboratory. A high-yield mutant strain Y19-11, BC yield reached 15.6g/L, was obtained by mutagenesis, screening and continuous passage. The mutant strain had excellent genetic stability and its BC yield increased by 44.4%,higher than the original strain. Meanwhile,through comparing the cell weight and yield of BC under different temperature and initial pH value,the optimal temperature and initial pH of mutant strain Y19-11 were determined as 30℃ and 5.5,respectively.
In order to obtain high bacterial cellulose(BC)-producing strain,composite mutagenesis of ultraviolet and diethyl sulfate was used to Gluconacetobacter xylinus BC13 which preserved in laboratory. A high-yield mutant strain Y19-11, BC yield reached 15.6g/L, was obtained by mutagenesis, screening and continuous passage. The mutant strain had excellent genetic stability and its BC yield increased by 44.4%,higher than the original strain. Meanwhile,through comparing the cell weight and yield of BC under different temperature and initial pH value,the optimal temperature and initial pH of mutant strain Y19-11 were determined as 30℃ and 5.5,respectively.
引文
[1]Brown A J.On an acetic ferment which forms cellulose[J].Journal of Chemical Society,1886,49:432-439.
[2]曹海鹏,袁帅,赵昆,等.细菌纤维素高产菌株的筛选及鉴定[J].食品科学,2010,32(5):211-214.
[3]陆胜民,贾静静,杨颖.细菌纤维素发酵工艺与应用研究进展[J].食品与发酵科技,2011,47(1):27-31.
[4]Wang Yan,Luo Qingping,Peng Bihui,et al.A novel thermotropic liquid crystalline:Benzoylated bacterial cellulose[J].Carbohydrate Polymers,2008,74(4):875-879.
[5]王银存,李利军,马英辉,等.细菌纤维素生产及应用研究进展[J].中国酿造,2011,229(4):20-23.
[6]Peggy O S,Cannon R E.Acetobacter ylinum:An Inq uiry into Cellulose Biosynthesis[J].The Alnerican Biology Teacher,2000,62(6):442-444.
[7]Bae SQ,Sugano Y,Ohi K,et al.Features of bacterial cellulose synthesis in a mutant generated by disruption of the diguanylate cyclase 1 gene of Acetobacter xylinum BPR2001[J].Applied Microbiology and Bioteehnology,2004,65(3):315-322.
[8]张雯,齐香君,李彦军,等.醋酸杆菌Acetobacter xylinum pde基因敲除研究[J].陕西科技大学学报,2013(5):121-125.
[9]汤卫华,李飞,贾原媛,等.细菌纤维素高产菌株的诱变选育和发酵条件研究[J].现代食品科技,2009,25(9):1016-1019.
[10]邓毛程,吴亚丽,梁世中.超声波促进高纤椰果发酵的研究[J].食品与发酵工业,2008,34(3):62-64.
[11]汤卫华,马霞,贾原媛,等.木醋杆菌流加静置培养生产细菌纤维素[J].食品与发酵工业,2008,34(2):21-24.
[12]施巧琴,吴松刚.工业微生物育种学[M].北京:科学出版社,2003:l33-147.
[13]帖金鑫,林娟,李小平,等.耐酸耐胆盐嗜酸乳杆菌的紫外诱变选育[J].食品研究与开发,2013,34(17):111-115.
[14]李严军,廖杰琼,青文哲,等.产类黄酮链霉菌的紫外线诱变选育[J].农产品加工·学刊,2013(9b):1-3.
[15]古丽娜孜,库米拉,张晓燕,等.低温淀粉酶高产菌株的诱变选育[J].新疆农业大学学报,2013,36(4):288-292.
[16]万中义,吴顺清,朱海云,等.细菌纤维素高产菌株的诱变育种[J].湖北农业科学,2010,49(12):3067-3068.
[17]B V Mohite,K K Kamalja,S V Patil.Statistical optimization of culture conditions for enhanced bacterial cellulose production by gluconoacetobacter hansenii ncim 2529[J].Cellulose,2012,19(5):1655-1666.
[18]邵伟,乐超银,熊泽,等.醋酸杆菌合成细菌纤维素的发酵动力学研究[J].中国酿造,2005,151(10):26-28.
[19]齐香君,张雯,韩戌瑁,等.细菌纤维素生产菌株的动力学研究[J].食品科学,2005,26(12):65-67.
[20]马霞,察可文,王瑞明,等.细菌纤维素静态发酵生产的动力学模型的确定[J].河南工业大学学报:自然科学版,2005,26(1):36-38.
[21]Dehui Lin,Patricia Lopez-Sanchez,Rui Li,et al.Production of bacterial cellulose by Gluconacetobacter hansenii CGMCC3917 using only waste beer yeast as nutrient source[J].Bioresource Technology,2014,151:113-119.
[22]Fa’bio P Gomes,Nuno HCS Silva,Eliane Trovatti,et al.Production of bacterial cellulose by Gluconacetobacter sacchari using dry olive mill residue[J].Biomass and Bioenergy,2013,55:205-211.
[23]Jyh-Ming Wu,Ren-Han Liu.Thin stillage supplementation greatly enhances bacterial cellulose production by Gluconacetobacter xylinus[J].Carbohydrate Polymers,2012,90:116-121.
[2]曹海鹏,袁帅,赵昆,等.细菌纤维素高产菌株的筛选及鉴定[J].食品科学,2010,32(5):211-214.
[3]陆胜民,贾静静,杨颖.细菌纤维素发酵工艺与应用研究进展[J].食品与发酵科技,2011,47(1):27-31.
[4]Wang Yan,Luo Qingping,Peng Bihui,et al.A novel thermotropic liquid crystalline:Benzoylated bacterial cellulose[J].Carbohydrate Polymers,2008,74(4):875-879.
[5]王银存,李利军,马英辉,等.细菌纤维素生产及应用研究进展[J].中国酿造,2011,229(4):20-23.
[6]Peggy O S,Cannon R E.Acetobacter ylinum:An Inq uiry into Cellulose Biosynthesis[J].The Alnerican Biology Teacher,2000,62(6):442-444.
[7]Bae SQ,Sugano Y,Ohi K,et al.Features of bacterial cellulose synthesis in a mutant generated by disruption of the diguanylate cyclase 1 gene of Acetobacter xylinum BPR2001[J].Applied Microbiology and Bioteehnology,2004,65(3):315-322.
[8]张雯,齐香君,李彦军,等.醋酸杆菌Acetobacter xylinum pde基因敲除研究[J].陕西科技大学学报,2013(5):121-125.
[9]汤卫华,李飞,贾原媛,等.细菌纤维素高产菌株的诱变选育和发酵条件研究[J].现代食品科技,2009,25(9):1016-1019.
[10]邓毛程,吴亚丽,梁世中.超声波促进高纤椰果发酵的研究[J].食品与发酵工业,2008,34(3):62-64.
[11]汤卫华,马霞,贾原媛,等.木醋杆菌流加静置培养生产细菌纤维素[J].食品与发酵工业,2008,34(2):21-24.
[12]施巧琴,吴松刚.工业微生物育种学[M].北京:科学出版社,2003:l33-147.
[13]帖金鑫,林娟,李小平,等.耐酸耐胆盐嗜酸乳杆菌的紫外诱变选育[J].食品研究与开发,2013,34(17):111-115.
[14]李严军,廖杰琼,青文哲,等.产类黄酮链霉菌的紫外线诱变选育[J].农产品加工·学刊,2013(9b):1-3.
[15]古丽娜孜,库米拉,张晓燕,等.低温淀粉酶高产菌株的诱变选育[J].新疆农业大学学报,2013,36(4):288-292.
[16]万中义,吴顺清,朱海云,等.细菌纤维素高产菌株的诱变育种[J].湖北农业科学,2010,49(12):3067-3068.
[17]B V Mohite,K K Kamalja,S V Patil.Statistical optimization of culture conditions for enhanced bacterial cellulose production by gluconoacetobacter hansenii ncim 2529[J].Cellulose,2012,19(5):1655-1666.
[18]邵伟,乐超银,熊泽,等.醋酸杆菌合成细菌纤维素的发酵动力学研究[J].中国酿造,2005,151(10):26-28.
[19]齐香君,张雯,韩戌瑁,等.细菌纤维素生产菌株的动力学研究[J].食品科学,2005,26(12):65-67.
[20]马霞,察可文,王瑞明,等.细菌纤维素静态发酵生产的动力学模型的确定[J].河南工业大学学报:自然科学版,2005,26(1):36-38.
[21]Dehui Lin,Patricia Lopez-Sanchez,Rui Li,et al.Production of bacterial cellulose by Gluconacetobacter hansenii CGMCC3917 using only waste beer yeast as nutrient source[J].Bioresource Technology,2014,151:113-119.
[22]Fa’bio P Gomes,Nuno HCS Silva,Eliane Trovatti,et al.Production of bacterial cellulose by Gluconacetobacter sacchari using dry olive mill residue[J].Biomass and Bioenergy,2013,55:205-211.
[23]Jyh-Ming Wu,Ren-Han Liu.Thin stillage supplementation greatly enhances bacterial cellulose production by Gluconacetobacter xylinus[J].Carbohydrate Polymers,2012,90:116-121.