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( Chao Gao ),( Min Jin ),( Zhiwei Yi ),( Runying Zeng ) 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.11
A novel sulfatase gene, ary423 (1,536 bp ORF), encoding a protein of 511 amino acids with a calculated molecular mass of 56 kDa, was identified from Flammeovirga pacifica, which was isolated from deep-sea sediments of west Pacific Ocean. Amino acid sequence analysis revealed that Ary423 possessed a conserved C-X-A-X-R motif, which was recognized as the sulfatase signature. Phylogenetic analysis suggested that Ary423 belonged to arylsulfatases. After heterologous expression in Escherichia coli cells, the recombinant Ary423 was purified with a N i+ affinity column, and was shown to be highly active at a broad range of temperatures from 30° to 70°C, with maximum activity at 40°C. Furthermore, recombinant Ary423 retained more than 70% and 40% of its maximum activity after 12 h of incubation at 50°C and 60°C, respectively, exhibiting good thermostability at high temperatures. The optimal pH for Ary423 was determined to be 8.0 and the activity of Ary423 could be slightly enhanced by Mg2+. The recombinant enzyme could hydrolyze sulfate ester bonds in pnitrophenyl sulfate (NPS) and Asparagus crude polysaccharides with a specific activity of 64.8 U/mg and 25.4 U/mg, respectively. These favorable properties could make Ary423 attractive for application in the desulfating process of agar production.
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Zhuhua Chan,Tianhua Zhong,Zhiwei Yi,Jing Xiao,Runying Zeng 한국생물공학회 2015 Biotechnology and Bioprocess Engineering Vol.20 No.5
The aim of this work is to enhance enduracidin production by Streptomyces sp. MC079. Based on the time course analysis of the specific cell growth rate and specific enduracidin formation rate, a two–stage pH control strategy was proposed to improve enduracidin production by shifting the culture pH from 5.5 to 5.8 after 112 h of cultivation. By applying this pH control strategy, enduracidin concentration and productivity was 51.2 and 65.0% higher than results with uncontrolled pH batch fermentation. For further enhancement of enduracidin production, the effects of constant–rate feeding and pH-shift feeding strategy were investigated. The results indicated that the pH-shift feeding strategy increased the maximum concentration and productivity of enduracidin to 61.37 mg/L and 0.697 mg/L/h in the constant–rate feeding fermentation process. This is 73.3 and 88.9% higher than results with uncontrolled pH batch fermentation, respectively. The obtained optimal pH shift feeding strategy may be useful for the industrial–scale microbial production of enduracidin.
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