Overexpression of an optimized Aspergillus sulphureus β-mannanase gene in Pichia pastoris

Chen, Xiaoling; Qiao, Jiaoyun; Yu, Haiqin; Cao, Yunhe

doi:10.2478/s11756-009-0043-5

Cited by 14 publications

(3 citation statements)

References 14 publications

(17 reference statements)

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“…During the past few years, numerous β-mannanase’s have been purified and characterized from various organisms including bacteria, yeasts, fungi and plants ( Cai et al, 2011 ). Our laboratory has expended a considerable amount of effort aimed at the development of β-mannanase’s for the food, pulp, and paper and oil drilling industries ( Chen et al, 2007 ; 2008 ; 2009 ; Li et al, 2010 ; Qiao et al, 2010 ; Cai et al, 2011 ).…”

Section: Introductionmentioning

confidence: 99%

Effects of Supplementation of β-Mannanase in Corn-soybean Meal Diets on Performance and Nutrient Digestibility in Growing Pigs

Chen

Guo

et al. 2013

Asian Australas. J. Anim. Sci

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A total of 288 crossbred (Duroc×Landrace×Yorkshire) growing pigs were used in two experiments to investigate the effects of adding β-mannanase to corn-soybean meal-based diets on pig performance and apparent total tract digestibility (ATTD). Both experiments lasted 28 d and were split into two phases namely 1 to 14 days (phase 1) and 15 to 28 days (phase 2). In Exp. 1,144 pigs weighing 23.60±1.59 kg BW were assigned to one of four corn-soybean meal-based diets containing 0, 200, 400 or 600 U/kg β-mannanase. Increasing the level of β-mannanase increased weight gain (quadratic effect; p<0.01) and feed efficiency (linear and quadratic effect; p<0.01) during the second phase and the overall experiment. However, performance was unaffected (p>0.05) by treatment during phase 1. Increasing the amount of β-mannanase in the diet improved (linear and quadratic effect; p<0.05) the ATTD of CP, NDF, ADF, calcium, and phosphorus during both phases. Based on the results of Exp. 1, the optimal supplementation level was determined to be 400 U/kg and this was the level that was applied in Exp. 2. In Exp. 2, 144 pigs weighing 23.50±1.86 kg BW were fed diets containing 0 or 400 U/kg of β-mannanase and 3,250 or 3,400 kcal/kg digestible energy (DE) in a 2×2 factorial design. β-Mannanase supplementation increased (p<0.01) weight gain and feed efficiency while the higher energy content increased (p<0.01) feed intake and feed efficiency during both phases and overall. Increased energy content and β-mannanase supplementation both increased (p<0.05) the ATTD of DM, CP, NDF, ADF, phosphorus, and GE during both phases. There were no significant interactions between energy level and β-mannanase for any performance or digestibility parameter. In conclusion, the β-mannanase used in the present experiment improved the performance of growing pigs fed diets based on corn and soybean. The mechanism through which the improvements were obtained appears to be related to improvements in ATTD.

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Section: Introductionmentioning

confidence: 99%

Effects of Supplementation of β-Mannanase in Corn-soybean Meal Diets on Performance and Nutrient Digestibility in Growing Pigs

Chen

Guo

et al. 2013

Asian Australas. J. Anim. Sci

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“…After the effect of different concentrations of methanol in the BMMY medium on the recombinant A. niger β-mannanase production by the GSKZαM2 strain was examined, we found that 1.5% (v/v) methanol in the BMMY medium was most suitable for induction for the recombinant A. niger β-mannanase production and the optimal induction time was 7 days. Under optimal conditions, 222.8 U/mL of β-mannanase activity was produced, which was higher than that of β-mannanase expressed in P. pastoris by codon optimization and double vector system (Chen et al, 2009;Tang et al, 2014).…”

Section: Discussionmentioning

confidence: 88%

Enhancing the expression of Aspergillus niger β-mannanase in Pichia pastoris by coexpression of protein disulfide isomerase

Chen¹,

Zhou²,

Xu³

et al. 2015

Turk J Biol

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A gene encoding β-mannanase from Aspergillus niger GIM3.452 was amplified and inserted into a pPIC9K vector. The resulting recombinant plasmid, pPIC9K-MAN, was transformed into Pichia pastoris GS115. One strain (GSKM-1) having the highest β-mannanase activity of 26.6 U/mL was obtained. In order to increase the secretion of β-mannanase in P. pastoris, we constructed a double recombinant yeast and made it coexpress protein disulfide isomerase. One strain (GSKZαM2) with the highest β-mannanase activity of 40 U/mL was then obtained and used to optimize expression conditions. When the GSKZαM2 strain was induced under the optimized conditions (methanol concentration 1.5%, induction time 7 days), β-mannanase activity reached 222.8 U/mL. SDS-PAGE and deglycosylation assays demonstrated that the recombinant A. niger β-mannanase, a glycosylated protein with an apparent molecular weight of 45 kDa, was secreted into the culture medium. It displayed maximum activity at pH 4.4 and 60 °C, and it was stable in a pH range of 2.4-8.0 and at a temperature of 60 °C or below. Our results suggested that coexpression chaperones could improve the yield of β-mannanase.

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“…The bias of codon usage has been demonstrated to be related to gene expression level (Sharp & Li 1986). In our previous studies, expression level of enzymes, such as β-mananase and β-xylanase, could be significantly increased by codon optimization (Cao et al 2007;Chen et al 2009). In this study, the optimized gene glu-opt coded for B. subtilis MA139 β-1,3-1,4-glucanase was overexpressed with a yield of 15,000 U/mL in a 10-L fermentor.…”

Section: Characterization Of the Recombinant β-13-14-glucanasementioning

confidence: 99%

Codon optimization, expression and characterization of Bacillus subtilis MA139 β-1,3-1,4-glucanase in Pichia pastoris

et al. 2010

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β-1,3-1,4-Glucanase has been broadly used in feed and brewing industries. According to the codon bias of Pichia pastoris, the Bacillus subtilis MA139 β-1,3-1,4-glucanase gene was de novo synthesized and expressed in P. pastoris X-33 strain under the control of the alcohol oxidase 1 promoter. In a 10-L fermentor, the β-1,3-1,4-glucanase was overexpressed with a yield of 15,000 U/mL by methanol induction for 96 h. The recombinant β-1,3-1,4-glucanase exhibited optimal activity at 40• C and pH 6.4. The activity of the recombinant β-1,3-1,4-glucanase was not significantly affected by various metal ions and chemical reagents. To our knowledge, the expression of this β-1,3-1,4-glucanase from Bacillus sp. in P. pastoris is in relatively high level compared to previous reports. These biochemical characteristics suggest that the recombinant β-1,3-1,4-glucanase has a prospective application in feed and brewing industries.

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Overexpression of an optimized Aspergillus sulphureus β-mannanase gene in Pichia pastoris

Cited by 14 publications

References 14 publications

Effects of Supplementation of β-Mannanase in Corn-soybean Meal Diets on Performance and Nutrient Digestibility in Growing Pigs

Effects of Supplementation of β-Mannanase in Corn-soybean Meal Diets on Performance and Nutrient Digestibility in Growing Pigs

Enhancing the expression of Aspergillus niger β-mannanase in Pichia pastoris by coexpression of protein disulfide isomerase

Codon optimization, expression and characterization of Bacillus subtilis MA139 β-1,3-1,4-glucanase in Pichia pastoris

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