Disulfide bonds elimination of endoglucanase II from Trichoderma reesei by site-directed mutagenesis to improve enzyme activity and thermal stability: An experimental and theoretical approach
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Akbarzadeh, Ali
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NIGEB, Dept Plant Biotechnol, Tehran, Iran
Shahid Beheshti Univ Med Sci, Gene Transfer Pioneers Res Grp, Tehran, Iran
Shahid Beheshti Univ, Biotechnol Prot Res Ctr PRC, GC, Tehran, IranNIGEB, Dept Plant Biotechnol, Tehran, Iran
Endoglucanasell (Cel5A) of Trichoderma reesei is widely used industrially with the high catalytic efficiency, but it is not stable high temperatures. Structural comparison with the closest thermophilic endoglucanase homolog, Cel5A from Thermoascus aurantiacus, demonstrates disulfide bond differences. Replacement of Cysteine99 with Valine and Cysteine323 with Histidine by site directed mutagenesis caused elimination of two disulfide bonds. Recombinant expression in Pichia pastoris showed the catalytic efficiency (kcat/Km) increment toward CMC for single mutant enzymes, C99V and C323H, about 1.87 and 13 folded respectively. This indicates that the elimination of disulfide bond in substrate binding cleft around the catalytic domain of mutant Endoglucanasell may be increased the flexibility of protein, to form a suitable E-S complex. In direct contrast with previous studies suggesting the existence of disulfide bonds increase the protein stability, the results showed mutant endoglucanase enzymes with disulfide bond reduction have higher thermal stability. The thermal stability of C99V and C323H in 80 degrees C were increased 2.4 and 234 folded, respectively. In this project, theoretical data had a good agreement with the experimental results. Because of high enzyme activity and thermal stability, both of C99V and C323H mutant have high potential suitable for different industrial applications. (C) 2018 Published by Elsevier B.V.
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King Saud Univ, Dept Pharmaceut Chem, Coll Pharm, Riyadh 11451, Saudi ArabiaKing Saud Univ, Dept Pharmaceut Chem, Coll Pharm, Riyadh 11451, Saudi Arabia
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Semnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, Iran
Shahid Beheshti Univ Med Sci, Sch Pharm, Gene Transfer Pioneers GTP Res Grp, Vali Asr Ave, Tehran, IranSemnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, Iran
Aghaeepoor, Mojtaba
Kobarfard, Farzad
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Shahid Beheshti Univ Med Sci, Sch Pharm, Dept Med Chem, Vali Asr Ave, Tehran, IranSemnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, Iran
Kobarfard, Farzad
Eidgahi, Mohammad Reza Akbari
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Semnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, IranSemnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, Iran
Eidgahi, Mohammad Reza Akbari
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Shabani, Aliakbar
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Dehnavi, Ehsan
Ahmadi, Maryam
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Semnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, IranSemnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, Iran
Ahmadi, Maryam
Akbarzadeh, Ali
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Shahid Beheshti Univ Med Sci, Sch Pharm, Gene Transfer Pioneers GTP Res Grp, Vali Asr Ave, Tehran, IranSemnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, Iran
机构:
Aligarh Muslim Univ, Fac Life Sci, Dept Biochem, Aligarh, Uttar Pradesh, IndiaAligarh Muslim Univ, Fac Life Sci, Dept Biochem, Aligarh, Uttar Pradesh, India
Alam, Md. Maroof
Qais, Faizan Abul
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Aligarh Muslim Univ, Dept Agr Microbiol, Aligarh, Uttar Pradesh, IndiaAligarh Muslim Univ, Fac Life Sci, Dept Biochem, Aligarh, Uttar Pradesh, India
Qais, Faizan Abul
Ahmad, Iqbal
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Aligarh Muslim Univ, Dept Agr Microbiol, Aligarh, Uttar Pradesh, IndiaAligarh Muslim Univ, Fac Life Sci, Dept Biochem, Aligarh, Uttar Pradesh, India
Ahmad, Iqbal
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Alam, Parvez
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Khan, Rizwan Hasan
Naseem, Imrana
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Aligarh Muslim Univ, Fac Life Sci, Dept Biochem, Aligarh, Uttar Pradesh, IndiaAligarh Muslim Univ, Fac Life Sci, Dept Biochem, Aligarh, Uttar Pradesh, India
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Aligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, IndiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
Naseem, Farha
Abdelhameed, Ali Saber
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King Saud Univ, Coll Pharm, Dept Pharmaceut Chem, Riyadh 11451, Saudi ArabiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
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Aligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, IndiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
Chaturvedi, Sumit Kumar
Anwar, Tamanna
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Ctr Bioinformat Res & Technol, Aligarh 202002, Uttar Pradesh, IndiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
Anwar, Tamanna
Siddiqi, Mohammad Khursheed
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Aligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, IndiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
Siddiqi, Mohammad Khursheed
Ajmal, Mohd Rehan
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Aligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, IndiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
Ajmal, Mohd Rehan
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Badr, Gamal
Mahmoud, Mohamed H.
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Natl Res Ctr, Food Sci & Nutr Dept, Cairo, Egypt
King Saud Univ, Sci Res, Riyadh, Saudi ArabiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
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Aligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, IndiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
Rabbani, Gulam
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Badr, Gamal
Badr, Badr Mohamed
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NCRRT, Radiat Biol Dept, Cairo, EgyptAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
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King Saud Univ, Dept Pharmaceut Chem, Coll Pharm, Riyadh 11451, Saudi ArabiaKing Saud Univ, Dept Pharmaceut Chem, Coll Pharm, Riyadh 11451, Saudi Arabia
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Semnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, Iran
Shahid Beheshti Univ Med Sci, Sch Pharm, Gene Transfer Pioneers GTP Res Grp, Vali Asr Ave, Tehran, IranSemnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, Iran
Aghaeepoor, Mojtaba
Kobarfard, Farzad
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Shahid Beheshti Univ Med Sci, Sch Pharm, Dept Med Chem, Vali Asr Ave, Tehran, IranSemnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, Iran
Kobarfard, Farzad
Eidgahi, Mohammad Reza Akbari
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Semnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, IranSemnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, Iran
Eidgahi, Mohammad Reza Akbari
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Shabani, Aliakbar
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Dehnavi, Ehsan
Ahmadi, Maryam
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Semnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, IranSemnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, Iran
Ahmadi, Maryam
Akbarzadeh, Ali
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Shahid Beheshti Univ Med Sci, Sch Pharm, Gene Transfer Pioneers GTP Res Grp, Vali Asr Ave, Tehran, IranSemnan Univ Med Sci, Semnan Biotechnol Res Ctr, Basij Blvd, Semnan, Iran
机构:
Aligarh Muslim Univ, Fac Life Sci, Dept Biochem, Aligarh, Uttar Pradesh, IndiaAligarh Muslim Univ, Fac Life Sci, Dept Biochem, Aligarh, Uttar Pradesh, India
Alam, Md. Maroof
Qais, Faizan Abul
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h-index: 0
机构:
Aligarh Muslim Univ, Dept Agr Microbiol, Aligarh, Uttar Pradesh, IndiaAligarh Muslim Univ, Fac Life Sci, Dept Biochem, Aligarh, Uttar Pradesh, India
Qais, Faizan Abul
Ahmad, Iqbal
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Aligarh Muslim Univ, Dept Agr Microbiol, Aligarh, Uttar Pradesh, IndiaAligarh Muslim Univ, Fac Life Sci, Dept Biochem, Aligarh, Uttar Pradesh, India
Ahmad, Iqbal
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Alam, Parvez
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Khan, Rizwan Hasan
Naseem, Imrana
论文数: 0引用数: 0
h-index: 0
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Aligarh Muslim Univ, Fac Life Sci, Dept Biochem, Aligarh, Uttar Pradesh, IndiaAligarh Muslim Univ, Fac Life Sci, Dept Biochem, Aligarh, Uttar Pradesh, India
机构:
Aligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, IndiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
Naseem, Farha
Abdelhameed, Ali Saber
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h-index: 0
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King Saud Univ, Coll Pharm, Dept Pharmaceut Chem, Riyadh 11451, Saudi ArabiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
机构:
Aligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, IndiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
Chaturvedi, Sumit Kumar
Anwar, Tamanna
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h-index: 0
机构:
Ctr Bioinformat Res & Technol, Aligarh 202002, Uttar Pradesh, IndiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
Anwar, Tamanna
Siddiqi, Mohammad Khursheed
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h-index: 0
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Aligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, IndiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
Siddiqi, Mohammad Khursheed
Ajmal, Mohd Rehan
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Aligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, IndiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
Ajmal, Mohd Rehan
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Badr, Gamal
Mahmoud, Mohamed H.
论文数: 0引用数: 0
h-index: 0
机构:
Natl Res Ctr, Food Sci & Nutr Dept, Cairo, Egypt
King Saud Univ, Sci Res, Riyadh, Saudi ArabiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
机构:
Aligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, IndiaAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
Rabbani, Gulam
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机构:
Badr, Gamal
Badr, Badr Mohamed
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NCRRT, Radiat Biol Dept, Cairo, EgyptAligarh Muslim Univ, Interdisciplinary Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India