可得然胶生产菌种的筛选及发酵条件优化
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摘要
采用苯胺蓝平板筛选法从土壤中筛选并鉴定了一株产可得然胶的根瘤菌,并对该菌株发酵合成可得然胶的培养基进行优化。在单因素优化的基础上,设计4因素3水平的正交优化方案,优化结果显示,该菌株发酵产可得然胶的最优培养基组成为:葡萄糖50 g/L,(NH4)2HPO42g/L,KH2PO41.5 g/L,Mg SO41.25 g/L,Fe SO4·7H2O 0.05 g/L,Mn SO40.02 g/L,Co Cl2·6H2O 0.01 g/L,Zn Cl20.01 g/L。经摇瓶发酵验证,可得然胶的产量和糖转化率较优化前分别提高了29.2%和5.4%,为进一步利用该根瘤菌发酵合成可得然胶奠定了良好基础。
A Rhizobium sp. capable of producing curdlan was isolated from soil. The polysaccharide product was identified to be curdlan by testing the thermal property,molecule composition,infrared spectrum and molecule weight. Furthermore,the orthogonal optimization experiment was conducted to optimize the curdlan fermentation medium,and the best medium composition was 50 g/L glucose,2 g/L(NH4)2 HPO4,1.5 g/L KH2 PO4,1.25 g/L Mg SO4,0.05 g/L Fe SO4·7 H2 O,0.02 g/L Mn SO4,0.01 g/L Co Cl2·6 H2 O,0.01 g/L Zn Cl2. Consequently,the curdlan production and glucose conversion rate were enhanced by 29.2% and 5.4% compared with those before optimization.
A Rhizobium sp. capable of producing curdlan was isolated from soil. The polysaccharide product was identified to be curdlan by testing the thermal property,molecule composition,infrared spectrum and molecule weight. Furthermore,the orthogonal optimization experiment was conducted to optimize the curdlan fermentation medium,and the best medium composition was 50 g/L glucose,2 g/L(NH4)2 HPO4,1.5 g/L KH2 PO4,1.25 g/L Mg SO4,0.05 g/L Fe SO4·7 H2 O,0.02 g/L Mn SO4,0.01 g/L Co Cl2·6 H2 O,0.01 g/L Zn Cl2. Consequently,the curdlan production and glucose conversion rate were enhanced by 29.2% and 5.4% compared with those before optimization.
引文
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[6]ZHANG Hongtao,ZHAN Xiaobei,ZHENG Zhiyong,et al.Advances in curdlan of production by Alcaligenes faecalis and function of curdlan[J].Chinese Journal of Bioprocess Engineering,2009,7(5):8-12.(in Chinese)
[7]ZHANG R,EDGAR K J.Properties,chemistry,and applications of the bioactive polysaccharide curdlan[J].Biomacromolecules,2014,15(4):1079-1096.
[8]LEHTOVAARA B C,GU F X.Pharmacological,structural,and drug delivery properties and applications of 1,3-β-glucans[J].Journal of Agricultural and Food Chemistry,2011,59(13):6813-6828.
[9]BAKER G C,SMITH J J,COWAN D A.Review and reanalysis of domain-specific 16S primer[J].Journal of Microbiological Methods,2003,55(3):541-555.
[10]GB28304-2012,食品安全国家标准食品添加剂可得然胶[S].北京:中国标准出版社,2012.
[11]KALYANASUNDARAM G T,DOBLE M,GUMMADI S N.2012.Production and downstream processing of(1-3)-beta-D-glucan from mutant strain of Agrobacterium sp.ATCC 31750[J].AMB Express,2(1):31.
[12]MILLER G L.Use of dinitrosalicylic acid reagent for determination of reducing sugar[J].Analytical Chemistry,1959,31(3):426-428.
[13]HARWOOD J,HUYSER D.Automated analysis of ammonia in water[J].Water Research,1970,4(10):695-704.
[14]TENG Jitao,CUI Jiandong.Screening and identification of a bacterial strain produced by curdlan and resarch on its culture conditions[J].China Condiment,2013,38(8):113-116.(in Chinese)
[15]LIU Y,GU Q,OFOSU F K,et al.Isolation and characterization of curdlan produced by Agrobacterium HX1126 using alpha-lactose as substrate[J].International Journal of Biological Macromolecules,2015,81:498-503.
[16]YU L,WU J,LIU J,et al.Enhanced curdlan production in Agrobacterium sp.ATCC 31749 by addition of low-polyphosphates[J].Biotechnology and Bioprocess Engineering,2011,16(1):34-41.
[17]WU J,ZHAN X,LIU H,et al.Enhanced production of curdlan by Alcaligenes faecalis by selective feeding with ammonia water during the cell growth phase of fermentation[J].Chinese Journal of Biotechnology,2008,24(6):1035-1039.
[18]NAKATA M,KAWAGUCHI T,KODAMA Y,et al.Characterization of curdlan in aqueous sodium hydroxide[J].Polymer,1998,39(6):1475-1481.
[19]KIM M K,RYU K E,CHOI W A,et al.Enhanced production of(1,3)-β-D-glucan by a mutant strain of Agrobacterium species[J].Biochemical Engineering Journal,2003,16(2):163-168.
[20]MARTINEZ C O,RUIZ S P,NOGUEIRA M T,et al.Effective immobilization of Agrobacterium sp.IFO 13140 cells in loofa sponge for curdlan biosynthesis[J].Molecules,2015,20(5):7957-7973.
[2]GHAI S K,HISAMATSU M,AMEMURA A,et al.Production and chemical composition of extracellular polysaccharides of Rhizobium[J].Journal of General Microbiology,1981,122:33-40.
[3]NAKANISHI I,KIMURA K,SUZUKI T,et al.Demonstration of curdlan-type polysaccharide and otherβ-1,3-glucan in microorganisms with aniline blue[J].The Journal of General and Applied Microbiology,1976,22:1-11.
[4]ZHANG H,NISHINARI K,WILLIAMS M A K,et al.A molecular description of the gelation mechanism of curdlan[J].International Journal of Biological Macromolecules,2002,30(1):7-16.
[5]LEIGH J A,DODSWORTH J A.Nitrogen regulation in bacteria and archaea[J].Annual Review of Microbiology,2007,61:349-377.
[6]ZHANG Hongtao,ZHAN Xiaobei,ZHENG Zhiyong,et al.Advances in curdlan of production by Alcaligenes faecalis and function of curdlan[J].Chinese Journal of Bioprocess Engineering,2009,7(5):8-12.(in Chinese)
[7]ZHANG R,EDGAR K J.Properties,chemistry,and applications of the bioactive polysaccharide curdlan[J].Biomacromolecules,2014,15(4):1079-1096.
[8]LEHTOVAARA B C,GU F X.Pharmacological,structural,and drug delivery properties and applications of 1,3-β-glucans[J].Journal of Agricultural and Food Chemistry,2011,59(13):6813-6828.
[9]BAKER G C,SMITH J J,COWAN D A.Review and reanalysis of domain-specific 16S primer[J].Journal of Microbiological Methods,2003,55(3):541-555.
[10]GB28304-2012,食品安全国家标准食品添加剂可得然胶[S].北京:中国标准出版社,2012.
[11]KALYANASUNDARAM G T,DOBLE M,GUMMADI S N.2012.Production and downstream processing of(1-3)-beta-D-glucan from mutant strain of Agrobacterium sp.ATCC 31750[J].AMB Express,2(1):31.
[12]MILLER G L.Use of dinitrosalicylic acid reagent for determination of reducing sugar[J].Analytical Chemistry,1959,31(3):426-428.
[13]HARWOOD J,HUYSER D.Automated analysis of ammonia in water[J].Water Research,1970,4(10):695-704.
[14]TENG Jitao,CUI Jiandong.Screening and identification of a bacterial strain produced by curdlan and resarch on its culture conditions[J].China Condiment,2013,38(8):113-116.(in Chinese)
[15]LIU Y,GU Q,OFOSU F K,et al.Isolation and characterization of curdlan produced by Agrobacterium HX1126 using alpha-lactose as substrate[J].International Journal of Biological Macromolecules,2015,81:498-503.
[16]YU L,WU J,LIU J,et al.Enhanced curdlan production in Agrobacterium sp.ATCC 31749 by addition of low-polyphosphates[J].Biotechnology and Bioprocess Engineering,2011,16(1):34-41.
[17]WU J,ZHAN X,LIU H,et al.Enhanced production of curdlan by Alcaligenes faecalis by selective feeding with ammonia water during the cell growth phase of fermentation[J].Chinese Journal of Biotechnology,2008,24(6):1035-1039.
[18]NAKATA M,KAWAGUCHI T,KODAMA Y,et al.Characterization of curdlan in aqueous sodium hydroxide[J].Polymer,1998,39(6):1475-1481.
[19]KIM M K,RYU K E,CHOI W A,et al.Enhanced production of(1,3)-β-D-glucan by a mutant strain of Agrobacterium species[J].Biochemical Engineering Journal,2003,16(2):163-168.
[20]MARTINEZ C O,RUIZ S P,NOGUEIRA M T,et al.Effective immobilization of Agrobacterium sp.IFO 13140 cells in loofa sponge for curdlan biosynthesis[J].Molecules,2015,20(5):7957-7973.