Cloning, overexpression and characterization of a thermostable pullulanase from Thermus thermophilus HB27

Wu, Huawei; Yu, Xin-Xin; Chen, Libing; Wu, Guangxu

doi:10.1016/j.pep.2013.11.010

Cited by 29 publications

(12 citation statements)

References 15 publications

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“…Recombinant expression is an essentially universal approach to obtaining high yields of pullulanase. Several host strains have been explored, including Escherichia coli [6,24,34], Bacillus subtilis [19,29,33], Bacillus licheniformis [1], Bacillus flavothermus [27], Pichia pastoris [35], Saccharomyces cerevisiae [17], and Raoultella planticola [13]. All of these hosts, except E. coli, showed poor expression of pullulanase.…”

Section: Introductionmentioning

confidence: 99%

Efficient extracellular expression of Bacillus deramificans pullulanase in Brevibacillus choshinensis

Zou

Duan

2016

Journal of Industrial Microbiology and Biotechnology

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In this study, the pullulanase gene from Bacillus deramificans was efficiently expressed in Brevibacillus choshinensis. The optimal medium for protein expression was determined through a combination of single-factor experiments and response surface methodology. The initial pH of the medium and the culture temperature were optimized. The pullulanase yield increased 10.8-fold through medium and condition optimization at the shake-flask level. From the results of these experiments, the dissolved oxygen level was optimized in a 3-L fermentor. Under these optimized conditions, the pullulanase activity and the specific pullulanase productivity reached 1005.8 U/mL and 110.5 × 10(3) U/g dry cell weight, respectively, with negligible intracellular expression. The Brevibacillus choshinensis expression system has proven to be valuable for the extracellular production of pullulanase.

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Section: Introductionmentioning

confidence: 99%

Efficient extracellular expression of Bacillus deramificans pullulanase in Brevibacillus choshinensis

Zou

Duan

2016

Journal of Industrial Microbiology and Biotechnology

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“…Interestingly, TTHpu produced a mixture of maltotriose, maltose and glucose as products from polysaccharide degradation, while most pullulanases usually form maltose and smaller oligosaccharides as main products [9]. However, a major problem for the efficient production of heat-stable amylopullulanases is their immense size, which has often been described to be up to 224 kDa.…”

Section: Current Opinion In Microbiologymentioning

confidence: 99%

Exploration of extremophiles for high temperature biotechnological processes

Elleuche

Schäfers

Blank

et al. 2015

Current Opinion in Microbiology

117

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“…To solve these problems, researchers have isolated new types of pullulanase enzymes from various microorganisms, such as Bacillus sp. AV-7 (Kunamneni and Singh, 2006 ), Thermus thermophiles (Wu et al, 2014 ), Bacillus deramificans (Duan et al, 2013 ), Anoxybacillus sp. SK3-4 (Kahar et al, 2016 ), and Bacillus naganoensis .…”

Section: Introductionmentioning

confidence: 99%

Enhanced Expression of Pullulanase in Bacillus subtilis by New Strong Promoters Mined From Transcriptome Data, Both Alone and in Combination

et al. 2018

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Pullulanase plays an important role as a starch hydrolysis enzyme in the production of bio-fuels and animal feed, and in the food industry. Compared to the methods currently used for pullulanase production, synthesis by Bacillus subtilis would be safer and easier. However, the current yield of pullulanase from B. subtilis is low to meet industrial requirements. Therefore, it is necessary to improve the yield of pullulanase by B. subtilis. In this study, we mined 10 highly active promoters from B. subtilis based on transcriptome and bioinformatic data. Individual promoters and combinations of promoters were used to improve the yield of pullulanase in B. subtilis BS001. Four recombinant strains with new promoters (Phag, PtufA, PsodA, and PfusA) had higher enzyme activity than the control (PamyE). The strain containing PsodA+fusA (163 U/mL) and the strain containing PsodA+fusA+amyE (336 U/mL) had the highest activity among the analyzed dual- and triple-promoter construct stains in shake flask, which were 2.29 and 4.73 times higher than that of the strain with PamyE, respectively. Moreover, the activity of the strain containing PsodA+fusA+amyE showed a maximum activity of 1,555 U/mL, which was 21.9 times higher than that of the flask-grown PamyE strain in a 50-liter fermenter. Our work showed that these four strong promoters mined from transcriptome data and their combinations could reliably increase the yield of pullulanase in quantities suitable for industrial applications.

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Cloning, overexpression and characterization of a thermostable pullulanase from Thermus thermophilus HB27

Cited by 29 publications

References 15 publications

Efficient extracellular expression of Bacillus deramificans pullulanase in Brevibacillus choshinensis

Efficient extracellular expression of Bacillus deramificans pullulanase in Brevibacillus choshinensis

Exploration of extremophiles for high temperature biotechnological processes

Enhanced Expression of Pullulanase in Bacillus subtilis by New Strong Promoters Mined From Transcriptome Data, Both Alone and in Combination

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