Optimized expression, purification and characterization of a family 11 xylanase (<scp>AuXyn11A</scp>) from <i>Aspergillus usamii</i> E001 in <i>Pichia pastoris</i>

Zhang, Huimin; Wang, Junqing; Wu, Minchen; Gao, Siyu; Li, Jianfang; Yang, Yongjian

doi:10.1002/jsfa.6309

Cited by 7 publications

(4 citation statements)

References 28 publications

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“…In addition, the pH condition is also an important factor for optimizing the expression of yeast recombinant proteins . It was reported that lowering the pH during the production stage generally mitigated protease activity and improved product quality . However, Fig.…”

Section: Resultsmentioning

confidence: 99%

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High‐level extracellular expression of inulin fructotransferase in Pichia pastoris for DFA III production

Zhan

Jiang

et al. 2014

J Sci Food Agric

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Section: Resultsmentioning

confidence: 99%

“…27 It was reported that lowering the pH during the production stage generally mitigated protease activity and improved product quality. 28 However, Fig. 2C shows that a relatively high pH value of 6.5 was the most suitable pH condition for IFTase expression by GS115-IFTase.…”

Section: Effect Of Culture Conditions On Recombinant Iftase Productionmentioning

confidence: 90%

High‐level extracellular expression of inulin fructotransferase in Pichia pastoris for DFA III production

Zhan

Jiang

et al. 2014

J Sci Food Agric

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“…The activities of recombinant SyS1 and SyGDH expressed in two recombinant E. coli strains were assayed and compared. Finally, the reaction conditions for asymmetric reduction of m-CPC to (R)-CCE catalyzed by whole E. coli/Sygdh-Sys1 cells were optimized using a 'one-parameter-at-a-time' method (Zhang et al 2014). To our knowledge, this is the first report on the asymmetric reduction of m-CPC coupled with regeneration of NADPH in situ by recombinant E. coli cells co-expressing both SyS1 and SyGDH.…”

Section: Introductionmentioning

confidence: 99%

Reduction of m-chlorophenacyl chloride coupled with regeneration of NADPH by recombinant Escherichia coli cells co-expressing both carbonyl reductase and glucose 1-dehydrogenase

Zhu

et al. 2015

Ann Microbiol

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Both Sys1 and Sygdh, two codon-optimized genes encoding SyS1 and SyGDH, were synthesized based on the carbonyl reductase (S1) and glucose 1-dehydrogenase (GDH) gene sequences, respectively, from Candida magnoliae and Thermoplasma acidophilum, and co-expressed in Escherichia coli BL21(DE3) using two strategies. One strategy involved a recombinant E. coli strain (E. coli/Sygdh-Sys1) constructed by transforming a recombinant plasmid, pETDuet-Sygdh-Sys1, into E. coli BL21. The other strategy involved another recombinant E. coli strain (E. coli/Sys1/ Sygdh) obtained by co-transforming the recombinant plasmids pET-22b-Sys1 and pET-28a-Sygdh into E. coli BL21. The enzyme activity assays indicated that the activities of recombinant SyS1 and SyGDH (3.7 and 56.3 U/g wet cells) expressed in E. coli/Sygdh-Sys1 were higher than those (2.8 and 44.1 U/g wet cells) in E. coli/Sys1/Sygdh. Accordingly, E. coli/Sygdh-Sys1 was chosen, and its whole cells were used as catalysts for the asymmetric reduction of m-chlorophenacyl chloride (m-CPC) to the corresponding (R)-2-chloro-1-(3-chlorophenyl)ethanol [(R)-CCE] coupled with the regeneration of NADPH in situ. Under the optimized reaction conditions of 30 mM m-CPC, 50 mg/ml wet cells, 40 mM glucose and 0.2 mM NADP + at pH 7.0 and 35°C for 3 h, (R)-CCE was obtained with a molar yield of 99.2 % and an enantiomeric excess (e.e.) value of more than 99 %.

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“…In previous studies, an endo- β -1,4-xylanase with high specific activity and good enzymatic properties was isolated from A. usamii . Furthermore, the gene encoding endo- β -1,4-xylanase from A. usamii was cloned and expressed in P. pastoris [ 9 , 10 ]. However, the low expression level does not allow the recombinant xylanase to be applied practically and economically in industry.…”

Section: Introductionmentioning

confidence: 99%

Improved Production ofAspergillus usamiiendo-β-1,4-Xylanase inPichia pastorisvia Combined Strategies

Wang¹,

Li²,

Liu³

2016

BioMed Research International

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A series of strategies were applied to improve expression level of recombinant endo-β-1,4-xylanase from Aspergillus usamii (A. usamii) in Pichia pastoris (P. pastoris). Firstly, the endo-β-1,4-xylanase (xynB) gene from A. usamii was optimized for P. pastoris and expressed in P. pastoris. The maximum xylanase activity of optimized (xynB-opt) gene was 33500 U/mL after methanol induction for 144 h in 50 L bioreactor, which was 59% higher than that by wild-type (xynB) gene. To further increase the expression of xynB-opt, the Vitreoscilla hemoglobin (VHb) gene was transformed to the recombinant strain containing xynB-opt. The results showed that recombinant strain harboring the xynB-opt and VHb (named X33/xynB-opt-VHb) displayed higher biomass, cell viability, and xylanase activity. The maximum xylanase activity of X33/xynB-opt-VHb in 50 L bioreactor was 45225 U/mL, which was 35% and 115% higher than that by optimized (xynB-opt) gene and wild-type (xynB) gene. Finally, the induction temperature of X33/xynB-opt-VHb was optimized in 50 L bioreactor. The maximum xylanase activity of X33/xynB-opt-VHb reached 58792 U/mL when the induction temperature was 22°C. The results presented here will greatly contribute to improving the production of recombinant proteins in P. pastoris.

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Optimized expression, purification and characterization of a family 11 xylanase (AuXyn11A) from Aspergillus usamii E001 in Pichia pastoris

Abstract: Its high specific activity and good enzymatic properties suggest that reAuXyn11A is a potential candidate for applications in industrial processes.

Cited by 7 publications

References 28 publications

High‐level extracellular expression of inulin fructotransferase in Pichia pastoris for DFA III production

High‐level extracellular expression of inulin fructotransferase in Pichia pastoris for DFA III production

Reduction of m-chlorophenacyl chloride coupled with regeneration of NADPH by recombinant Escherichia coli cells co-expressing both carbonyl reductase and glucose 1-dehydrogenase

Improved Production ofAspergillus usamiiendo-β-1,4-Xylanase inPichia pastorisvia Combined Strategies

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