钝齿棒杆菌N-乙酰鸟氨酸转氨酶的克隆表达分析及其重组菌的精氨酸发酵
Cloning, expression and characterization of N-Acetylornithine aminotransferase from Corynebacterium crenatum and its effects on L-arginine fermentation
投稿时间:2010-12-14  
DOI:  
中文关键词:钝齿棒杆菌,L-精氨酸,乙酰鸟氨酸转氨酶,酶学性质,发酵
英文关键词:Corynebacterium crenatum, L-arginine, N-Acetylornithine aminotransferase, enzymatic properties, fermentation
基金项目:国家高技术研究发展计划 (863计划) (No. 2007AA02Z207),国家重点基础研究发展计划 (973计划) (No. 2007CB707804),教育部新世纪优秀人才计划 (Nos. NCET-07-0380, NCET-10-0459),国家自然科学基金 (No. 30970056),中央高校基本科研业务费专项资金 (No. JUSRP31001) 资助。
作者单位E-mail
徐美娟 江南大学 工业生物技术教育部重点实验室无锡 214122  
张显 江南大学 工业生物技术教育部重点实验室无锡 214122  
饶志明 江南大学 工业生物技术教育部重点实验室无锡 214122 raozm@yahoo.com.cn 
杨娟 江南大学 工业生物技术教育部重点实验室无锡 214122  
窦文芳 江南大学医药学院无锡 214122  
金坚 江南大学医药学院无锡 214122  
许正宏 江南大学医药学院无锡 214122 zhenghongxu@jiangnan.edu.cn 
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中文摘要:
      N-乙酰鸟氨酸转氨酶 (EC 2.6.1.11,ACOAT) 是钝齿棒杆菌Corynebacterium crenatum精氨酸合成途径中的第4个酶,催化底物N-乙酰谷氨酸半醛生成产物N-乙酰鸟氨酸。为研究N-乙酰鸟氨酸转氨酶在钝齿棒杆菌中精氨酸合成中的作用,考察其酶学性质,对培养基成分和发酵过程工艺条件的优化提高精氨酸产量提供依据。从精氨酸高产菌株钝齿棒杆菌SYPA 5-5染色体扩增获得ACOAT编码基因argD,全长1 176 bp,编码390个氨基酸,在Escherichia coli BL21(D
英文摘要:
      N-Acetylornithine aminotransferase (EC 2.6.1.11, ACOAT) catalyzes the conversion of N-acetylglutamic semialdehyde to N-acetylornithine, the forth step involved in the L-arginine biosynthetic pathways. We studied the enzyme properties to set up reliable theoretical basis for the arginine fermentation optimization. ACOAT encoding gene argD was cloned from an industrial L-arginine producer Corynebacterium crenatum SYPA 5-5. Analysis of argD sequences revealed that only one ORF existed, which coded a peptide of 390 amino acids with a calculated molecular weight of 41.0 kDa. The argD gene from C. crenatum SYPA 5-5 was expressed both in Escherichia coli BL21 and C. crenatum SYPA. Then ACOAT was purified by Ni-NTA affinity chromatography and its specific enzyme activity was 108.2 U/g. Subsequently, the expression plasmid pJCtac-CcargD was transformed into C. crenatum SYPA and the specific activity of ACOAT was improved evidently in the recombinant C. crenatum CCD. Further fermentative character of CCD1 was also analyzed. The results showed that the L-arginine producing ability of the recombinant strain was 39.7 g/L improved by 14.7%.
徐美娟,张显,饶志明,杨娟,窦文芳,金坚,许正宏.钝齿棒杆菌N-乙酰鸟氨酸转氨酶的克隆表达分析及其重组菌的精氨酸发酵[J].生物工程学报,2011,27(7):1013~1023
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