Hyperexpression of two Aspergillus niger xylanase genes in Escherichia coli and characterization of the gene products

Yi, Xiuli; Shi, Yan; Xu, Hui; Li, Wei; Xie, Jinlu; Yu, Rongqing; Zhu, Jun; Cao, Yi; Qiao, Dan

doi:10.1590/s1517-83822010000300030

Cited by 17 publications

(9 citation statements)

References 28 publications

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“…xylanase had a high homology to Ao_Xln_41 (99.9%) (Figure 1(b)). The xlnG2 (699 bp, 232 aa) from A. oryzae VTCC-F187 belongs to the glycosyl hydrolase family 11, which was completely similar to the xlnG2 (767 bp, 232 aa) (glycosyl hydrolase family 11) [14], A. oryzae xlnG1 (725 bp, 189 aa) [21], xlnC (636 bp, 211 aa) [22], xlnA (glycosyl hydrolase family 10) [16], and xlnD (957 bp, 318 aa) [23] from A. niger strains (from GenBank). The phylogenetic tree of nucleotide and amino acids are not much different; this can be explained by a multitude of codons encoding for an amino acid.…”

Section: Xylanase Gene Sequence Analysis After Nucleotide Sequencingmentioning

confidence: 84%

Cloning, Expression, and Characterization of Xylanase G2 from Aspergillus oryzae VTCC-F187 in Aspergillus niger VTCC-F017

Tuyến

Cuong

Thanh

et al. 2021

BioMed Research International

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The study focuses on engineering of recombinant Aspergillus niger to produce highly active xylanase. The xylanase G2 encoding gene originating from Aspergillus oryzae VTCC-F187 was cloned, amplified, and inserted into the pAN7.1GluA vector with specific primers possessing BamHI. The recombinant plasmid was introduced into Aspergillus niger VTCC-F017 by chemical methods. The recombinant strain was checked by polymerase chain reaction method and Southern blot. Next, the recombinant protein was expressed and purified by His-tag column. The molecular mass of the purified xylanase G2, as determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), was 21 kDa with a specific activity of 1025 IU/mg towards 0.5% (w/v) of birchwood xylan. The optimal temperature and pH were 55°C and pH 6.5, respectively. The enzyme was stable in a temperature ranges 25–40°C and a pH ranges 5–7. The presence of Tween 80 enhanced xylanase activity. Triton X-100, however, had no impact on the function of the enzyme. The xylanase activity was reduced by Tween 20, SDS, and organic solvents. The enzyme was completely inhibited by Hg2+ and partially by Zn2+, Fe2+, and Ag+, while it was slightly stimulated by K+ and EDTA.

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Section: Xylanase Gene Sequence Analysis After Nucleotide Sequencingmentioning

confidence: 84%

Cloning, Expression, and Characterization of Xylanase G2 from Aspergillus oryzae VTCC-F187 in Aspergillus niger VTCC-F017

Tuyến

Cuong

Thanh

et al. 2021

BioMed Research International

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“…The xylanase gene xynB (FJ772090)was cloned from A. niger SCTCC 400264 in the Sichuan Type Culture Collection Center (SCTCC, China) [26]. E. coli JM109 was used as a cloning host and DNA propagation, and was grown in LB medium.…”

Section: Strains Plasmids Reagents and Culturing Conditionsmentioning

confidence: 99%

Thermostable Sites and Catalytic Characterization of Xylanase XYNB of Aspergillus niger SCTCC 400264

Li¹,

Xu²,

Xie³

et al. 2014

Journal of Microbiology and Biotechnology

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In order to improve the expression of heat-resistant xylanase XYNB from Aspergillus niger SCTCC 400264, XynB has been cloned into Pichia pastoris secretary vector pPIC9K. The XynB production of recombinant P. pastoris was four times as E. coli, the Vmax and specific activity of XynB reached 2,547.7 μmol/mg and 4,757 U/mg, respectively. And the XynB still had 74% residual enzyme activity after 30 min-heat treatment at 80°C. The van der Waals force analysis in XYNB (ACN89393 and AAS67299), there is one more oxygen radicals in AAS67299 in their catalytic site, indicating that the local cavity is much more free, and it is more optimal for substrate binding, affinity reaction, and proton transfer etc, and eventually increasing enzyme activity. The H-bonds analysis of XYNB indicated that there are two more H-bonds in 33rd Ser of XYNB (AAS67299) than 33rd Ala(ACN89393 ), two H-bonds between Ser70 and Asp67.

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“…Other fungal xylanases have also been shown to be non-glycosylated enzymes [118]. In line with this observation, different groups have described the production of catalytically-active eukaryotic xylanases in E. coli [134, 135]. Thus, E. coli , although to a lesser extent due to their lack of secretion system, has also been used to study different bacterial xylanases [136, 137].…”

Section: Recombinant Fibrolytic Enzymesmentioning

confidence: 99%

Trends in recombinant protein use in animal production

et al. 2017

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Recombinant technologies have made possible the production of a broad catalogue of proteins of interest, including those used for animal production. The most widely studied proteins for the animal sector are those with an important role in reproduction, feed efficiency, and health. Nowadays, mammalian cells and fungi are the preferred choice for recombinant production of hormones for reproductive purposes and fibrolytic enzymes to enhance animal performance, respectively. However, the development of low-cost products is a priority, particularly in livestock. The study of cell factories such as yeast and bacteria has notably increased in the last decades to make the new developed reproductive hormones and fibrolytic enzymes a real alternative to the marketed ones. Important efforts have also been invested to developing new recombinant strategies for prevention and therapy, including passive immunization and modulation of the immune system. This offers the possibility to reduce the use of antibiotics by controlling physiological processes and improve the efficacy of preventing infections. Thus, nowadays different recombinant fibrolytic enzymes, hormones, and therapeutic molecules with optimized properties have been successfully produced through cost-effective processes using microbial cell factories. However, despite the important achievements for reducing protein production expenses, alternative strategies to further reduce these costs are still required. In this context, it is necessary to make a giant leap towards the use of novel strategies, such as nanotechnology, that combined with recombinant technology would make recombinant molecules affordable for animal industry.

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Hyperexpression of two Aspergillus niger xylanase genes in Escherichia coli and characterization of the gene products

Cited by 17 publications

References 28 publications

Cloning, Expression, and Characterization of Xylanase G2 from Aspergillus oryzae VTCC-F187 in Aspergillus niger VTCC-F017

Cloning, Expression, and Characterization of Xylanase G2 from Aspergillus oryzae VTCC-F187 in Aspergillus niger VTCC-F017

Thermostable Sites and Catalytic Characterization of Xylanase XYNB of Aspergillus niger SCTCC 400264

Trends in recombinant protein use in animal production

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