Catalysing the reversible interconversion of L-alanine and alpha-ketoglutarate to pyruvate and L-glutamate in the presence of pyridoxal 5 phosphate, alanine aminotransferase (ALT) is an enzyme which operates at the cross roads of amino acid and carbohydrate metabolism. The enzyme has been reported from a wide range of organisms including animals, plants, fungi and microbes. The enzyme has a clinical applications in the diagnosis of many diseases. In the present study we have produced a recombinant of ALT from Pyrococcus abyssi in BL21 (DE3) strain of E. coli. The recombinant enzyme was purified by anion exchange chromatography, it displayed a 45kDa band on SDS-PAGE, with 58.1% final recovery, 15.3 fold purification and 139 U/mg specific activity. Maximum enzyme activity was found at pH 8 and above 90C, its KM and Vmax values were found 25 mu M L-alanine and 149 U/min/mg respectively. In silico studies have shown that the enzyme was found in a monomer structure. Molecular docking studies with potential molecules involved in the reaction catalysed have been conducted and binding energy values were calculated for each molecule including L-alanine, pyridoxal 5 phosphate, pyruvate, alpha ketoglutarate and L-glutamate. Present study provides the first report of ALT from Pyrococcus abyssi and suggests active site residues of enzyme from archaeal origin.
机构:
Univ Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Duarte, Lovaine Silva
Barse, Laisa Quadros
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Pontificia Univ Catolica Rio Grande do Sul PUCRS, CPBMF, 92A TECNOPUC Bldg,4592 Av Bento Goncalves, BR-90650001 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Barse, Laisa Quadros
Dalberto, Pedro Ferrari
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Pontificia Univ Catolica Rio Grande do Sul PUCRS, CPBMF, 92A TECNOPUC Bldg,4592 Av Bento Goncalves, BR-90650001 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Dalberto, Pedro Ferrari
Santos da Silva, William Tadeu
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h-index: 0
机构:
Univ Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Santos da Silva, William Tadeu
Rodrigues, Rafael Costa
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h-index: 0
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Univ Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Rodrigues, Rafael Costa
Machado, Pablo
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机构:
Pontificia Univ Catolica Rio Grande do Sul PUCRS, CPBMF, 92A TECNOPUC Bldg,4592 Av Bento Goncalves, BR-90650001 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Machado, Pablo
Basso, Luiz Augusto
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h-index: 0
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Pontificia Univ Catolica Rio Grande do Sul PUCRS, CPBMF, 92A TECNOPUC Bldg,4592 Av Bento Goncalves, BR-90650001 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Basso, Luiz Augusto
Bizarro, Cristiano Valim
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h-index: 0
机构:
Pontificia Univ Catolica Rio Grande do Sul PUCRS, CPBMF, 92A TECNOPUC Bldg,4592 Av Bento Goncalves, BR-90650001 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Bizarro, Cristiano Valim
Zachia Ayub, Marco Antonio
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机构:
Univ Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
机构:
Shandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R ChinaShandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R China
Li, An-Na
Yu, Kai
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Shandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R ChinaShandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R China
Yu, Kai
Liu, Hai-Quan
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h-index: 0
机构:
Shandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R ChinaShandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R China
Liu, Hai-Quan
Zhang, Jie
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h-index: 0
机构:
Shandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R ChinaShandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R China
Zhang, Jie
Li, Hua
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机构:
Shandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R ChinaShandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R China
Li, Hua
Li, Duo-Chuan
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h-index: 0
机构:
Shandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R ChinaShandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R China
机构:
Chinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China
Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R ChinaChinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China
Zheng, Yingying
Xue, Yanfen
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h-index: 0
机构:
Chinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R ChinaChinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China
Xue, Yanfen
Zhang, Yueling
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h-index: 0
机构:
Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Tianjin 300308, Peoples R ChinaChinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China
Zhang, Yueling
Zhou, Cheng
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h-index: 0
机构:
Chinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R ChinaChinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China
Zhou, Cheng
Schwaneberg, Ulrich
论文数: 0引用数: 0
h-index: 0
机构:
Rhein Westfal TH Aachen, Lehrstuhl Biotechnol, D-52074 Aachen, GermanyChinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China
Schwaneberg, Ulrich
Ma, Yanhe
论文数: 0引用数: 0
h-index: 0
机构:
Chinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R ChinaChinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China
机构:
Univ Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Duarte, Lovaine Silva
Barse, Laisa Quadros
论文数: 0引用数: 0
h-index: 0
机构:
Pontificia Univ Catolica Rio Grande do Sul PUCRS, CPBMF, 92A TECNOPUC Bldg,4592 Av Bento Goncalves, BR-90650001 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Barse, Laisa Quadros
Dalberto, Pedro Ferrari
论文数: 0引用数: 0
h-index: 0
机构:
Pontificia Univ Catolica Rio Grande do Sul PUCRS, CPBMF, 92A TECNOPUC Bldg,4592 Av Bento Goncalves, BR-90650001 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Dalberto, Pedro Ferrari
Santos da Silva, William Tadeu
论文数: 0引用数: 0
h-index: 0
机构:
Univ Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Santos da Silva, William Tadeu
Rodrigues, Rafael Costa
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h-index: 0
机构:
Univ Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Rodrigues, Rafael Costa
Machado, Pablo
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h-index: 0
机构:
Pontificia Univ Catolica Rio Grande do Sul PUCRS, CPBMF, 92A TECNOPUC Bldg,4592 Av Bento Goncalves, BR-90650001 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Machado, Pablo
Basso, Luiz Augusto
论文数: 0引用数: 0
h-index: 0
机构:
Pontificia Univ Catolica Rio Grande do Sul PUCRS, CPBMF, 92A TECNOPUC Bldg,4592 Av Bento Goncalves, BR-90650001 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Basso, Luiz Augusto
Bizarro, Cristiano Valim
论文数: 0引用数: 0
h-index: 0
机构:
Pontificia Univ Catolica Rio Grande do Sul PUCRS, CPBMF, 92A TECNOPUC Bldg,4592 Av Bento Goncalves, BR-90650001 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
Bizarro, Cristiano Valim
Zachia Ayub, Marco Antonio
论文数: 0引用数: 0
h-index: 0
机构:
Univ Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, BrazilUniv Fed Rio Grande do Sul, Food Sci & Technol Inst, Biotechnol Bioproc & Biocatalysis Grp, Av Bento Goncalves 9500,POB 15090, BR-91501970 Porto Alegre, RS, Brazil
机构:
Shandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R ChinaShandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R China
Li, An-Na
Yu, Kai
论文数: 0引用数: 0
h-index: 0
机构:
Shandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R ChinaShandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R China
Yu, Kai
Liu, Hai-Quan
论文数: 0引用数: 0
h-index: 0
机构:
Shandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R ChinaShandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R China
Liu, Hai-Quan
Zhang, Jie
论文数: 0引用数: 0
h-index: 0
机构:
Shandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R ChinaShandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R China
Zhang, Jie
Li, Hua
论文数: 0引用数: 0
h-index: 0
机构:
Shandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R ChinaShandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R China
Li, Hua
Li, Duo-Chuan
论文数: 0引用数: 0
h-index: 0
机构:
Shandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R ChinaShandong Agr Univ, Dept Environm Biol, Tai An 271018, Shandong, Peoples R China
机构:
Chinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China
Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R ChinaChinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China
Zheng, Yingying
Xue, Yanfen
论文数: 0引用数: 0
h-index: 0
机构:
Chinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R ChinaChinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China
Xue, Yanfen
Zhang, Yueling
论文数: 0引用数: 0
h-index: 0
机构:
Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Tianjin 300308, Peoples R ChinaChinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China
Zhang, Yueling
Zhou, Cheng
论文数: 0引用数: 0
h-index: 0
机构:
Chinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R ChinaChinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China
Zhou, Cheng
Schwaneberg, Ulrich
论文数: 0引用数: 0
h-index: 0
机构:
Rhein Westfal TH Aachen, Lehrstuhl Biotechnol, D-52074 Aachen, GermanyChinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China
Schwaneberg, Ulrich
Ma, Yanhe
论文数: 0引用数: 0
h-index: 0
机构:
Chinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R ChinaChinese Acad Sci, State Key Lab Microbial Resources, Inst Microbiol, Beijing 100101, Peoples R China