Identification of archaeon-producing hyperthermophilic alpha-amylase and characterization of the alpha-amylase

doi:10.1007/s00253-008-1561-8

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	Identification of archaeon-producing hyperthermophilic alpha-amylase and characterization of the alpha-amylase
	Wang, Shujun 1,2; Lu, Zhaoxin 1; Lu, Mingsheng 2; Qin, Song 3; Liu, Hongfei 2; Deng, Xiangyuan 3; Lin, Qian 1; Chen, Jianan 2
	2008-09-01
发表期刊	APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
ISSN	0175-7598
卷号	80 期号:4 页码:605-614
文章类型	Article
摘要	The extremely thermophilic anaerobic archaeon strain, HJ21, was isolated from a deep-sea hydrothermal vent, could produce hyperthermophilic alpha-amylase, and later was identified as Thermococcus from morphological, biochemical, and physiological characteristics and the 16S ribosomal RNA gene sequence. The extracellular thermostable alpha-amylase produced by strain HJ21 exhibited maximal activity at pH 5.0. The enzyme was stable in a broad pH range from pH 5.0 to 9.0. The optimal temperature of alpha-amylase was observed at 95 degrees C. The half-life of the enzyme was 5 h at 90 degrees C. Over 40% and 30% of the enzyme activity remained after incubation at 100 degrees C for 2 and 3 h, respectively. The enzyme did not require Ca(2+) for thermostability. This alpha-amylase gene was cloned, and its nucleotide sequence displayed an open reading frame of 1,374 bp, which encodes a protein of 457 amino acids. Analysis of the deduced amino acid sequence revealed that four homologous regions common in amylases were conserved in the HJ21 alpha-amylase. The molecular weight of the mature enzyme was calculated to be 51.4 kDa, which correlated well with the size of the purified enzyme as shown by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis.; The extremely thermophilic anaerobic archaeon strain, HJ21, was isolated from a deep-sea hydrothermal vent, could produce hyperthermophilic alpha-amylase, and later was identified as Thermococcus from morphological, biochemical, and physiological characteristics and the 16S ribosomal RNA gene sequence. The extracellular thermostable alpha-amylase produced by strain HJ21 exhibited maximal activity at pH 5.0. The enzyme was stable in a broad pH range from pH 5.0 to 9.0. The optimal temperature of alpha-amylase was observed at 95 degrees C. The half-life of the enzyme was 5 h at 90 degrees C. Over 40% and 30% of the enzyme activity remained after incubation at 100 degrees C for 2 and 3 h, respectively. The enzyme did not require Ca2+ for thermostability. This alpha-amylase gene was cloned, and its nucleotide sequence displayed an open reading frame of 1,374 bp, which encodes a protein of 457 amino acids. Analysis of the deduced amino acid sequence revealed that four homologous regions common in amylases were conserved in the HJ21 alpha-amylase. The molecular weight of the mature enzyme was calculated to be 51.4 kDa, which correlated well with the size of the purified enzyme as shown by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
关键词	Hyperthermophilic Alpha-amylase Thermococcus Sp. Thermostability Archaeon Identification Characterization
学科领域	Biotechnology & Applied Microbiology
DOI	10.1007/s00253-008-1561-8
URL	查看原文
收录类别	SCI
语种	英语
WOS记录号	WOS:000258881400007
引用统计	被引频次：22[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/5664
专题	实验海洋生物学重点实验室
作者单位	1.Nanjing Agr Univ, Coll Food Sci & Technol, Nanjing 210095, Jiangsu, Peoples R China 2.Huaihai Inst Technol, Jiangsu Key Lab Marine Biotechnol, Lianyungang 222005, Jiangsu, Peoples R China 3.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Shandong, Peoples R China
推荐引用方式 GB/T 7714	Wang, Shujun,Lu, Zhaoxin,Lu, Mingsheng,et al. Identification of archaeon-producing hyperthermophilic alpha-amylase and characterization of the alpha-amylase[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2008,80(4):605-614.
APA	Wang, Shujun.,Lu, Zhaoxin.,Lu, Mingsheng.,Qin, Song.,Liu, Hongfei.,...&Chen, Jianan.(2008).Identification of archaeon-producing hyperthermophilic alpha-amylase and characterization of the alpha-amylase.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,80(4),605-614.
MLA	Wang, Shujun,et al."Identification of archaeon-producing hyperthermophilic alpha-amylase and characterization of the alpha-amylase".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 80.4(2008):605-614.

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