Improvement in the thermostability of a type A feruloyl esterase, AuFaeA, from Aspergillus usamii by iterative saturation mutagenesis

Verfasser / Beitragende:
[Xin Yin, Jian-Fang Li, Chun-Juan Wang, Die Hu, Qin Wu, Ying Gu, Min-Chen Wu]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/23(2015-12-01), 10047-10056
Format:
Artikel (online)
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024 7 0 |a 10.1007/s00253-015-6889-2  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-015-6889-2 
245 0 0 |a Improvement in the thermostability of a type A feruloyl esterase, AuFaeA, from Aspergillus usamii by iterative saturation mutagenesis  |h [Elektronische Daten]  |c [Xin Yin, Jian-Fang Li, Chun-Juan Wang, Die Hu, Qin Wu, Ying Gu, Min-Chen Wu] 
520 3 |a Feruloyl or ferulic acid esterase (Fae, EC 3.1.1.73) catalyzes the hydrolysis of ester bonds between polysaccharides and phenolic acid compounds in xylan side chain. In this study, the thermostability of a type A feruloyl esterase (AuFaeA) from Aspergillus usamii was increased by iterative saturation mutagenesis (ISM). Two amino acids, Ser33 and Asn92, were selected for saturation mutagenesis according to the B-factors analyzed by B-FITTER software and ΔΔG values predicted by PoPMuSiC algorithm. After screening the saturation mutagenesis libraries constructed in Pichia pastoris, 15 promising variants were obtained. The best variant S33E/N92-4 (S33E/N92R) produced a T m value of 44.5°C, the half-lives (t 1/2) of 35 and 198min at 55 and 50°C, respectively, corresponding to a 4.7°C, 2.33- and 3.96-fold improvement compared to the wild type. Additionally, the best S33 variant S33-6 (S33E) was thermostable at 50°C with a t 1/2 of 82min, which was 32min longer than that of the wild type. All the screened S33E/N92 variants were more thermostable than the best S33 variant S33-6 (S33E). This work would contribute to the further studies on higher thermostability modification of type A feruloyl esterases, especially those from fungi. The thermostable feruloyl esterase variants were expected to be potential candidates for industrial application in prompting the enzymic degradation of plant biomass materials at elevated temperatures. 
540 |a Springer-Verlag Berlin Heidelberg, 2015 
690 7 |a Feruloyl esterase  |2 nationallicence 
690 7 |a Aspergillus usamii  |2 nationallicence 
690 7 |a Thermostability  |2 nationallicence 
690 7 |a Iterative saturation mutagenesis  |2 nationallicence 
700 1 |a Yin  |D Xin  |u Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, China  |4 aut 
700 1 |a Li  |D Jian-Fang  |u State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, China  |4 aut 
700 1 |a Wang  |D Chun-Juan  |u State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, China  |4 aut 
700 1 |a Hu  |D Die  |u Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, China  |4 aut 
700 1 |a Wu  |D Qin  |u Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, China  |4 aut 
700 1 |a Gu  |D Ying  |u Wuxi Medical School, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, China  |4 aut 
700 1 |a Wu  |D Min-Chen  |u Wuxi Medical School, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, China  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/23(2015-12-01), 10047-10056  |x 0175-7598  |q 99:23<10047  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-015-6889-2  |q text/html  |z Onlinezugriff via DOI 
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900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
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950 |B NATIONALLICENCE  |P 700  |E 1-  |a Yin  |D Xin  |u Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Li  |D Jian-Fang  |u State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Wang  |D Chun-Juan  |u State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Hu  |D Die  |u Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Wu  |D Qin  |u Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Gu  |D Ying  |u Wuxi Medical School, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Wu  |D Min-Chen  |u Wuxi Medical School, Jiangnan University, 1800 Lihu Road, 214122, Wuxi, China  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/23(2015-12-01), 10047-10056  |x 0175-7598  |q 99:23<10047  |1 2015  |2 99  |o 253