Expression of recombinant Aspergillus niger xylanase A in Pichia pastoris and its action on xylan

Liu, Mingqi; Weng, Xiao-Yan; Sun, Jiazeng

doi:10.1016/j.pep.2006.04.007

Cited by 41 publications

(21 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These results confirm that xylanase PXII-1 is a xylan endo-acting enzyme. Purified endoxylanase from A. carneus [21] and A. niger [43] showed a similar product profile nevertheless only xylooligosaccharides were detected upon xylan hydrolysis by A. versicolor xylanase [12]. Enzymes with high endoxylanase activity and low exo-xylanase and/or b-xylosidase activity are favored for the production of xylooligosaccharides (XOs) [63] that can be used as prebiotics.…”

Section: Xylanase Hydrolytic Profilementioning

confidence: 99%

Purification and characterization studies of a thermostable β-xylanase from Aspergillus awamori

Teixeira

Siqueira

Souza

et al. 2010

J Ind Microbiol Biotechnol

View full text Add to dashboard Cite

This study presents data on the production, purification, and properties of a thermostable β-xylanase produced by an Aspergillus awamori 2B.361 U2/1 submerged culture using wheat bran as carbon source. Fractionation of the culture filtrate by membrane ultrafiltration followed by Sephacryl S-200 and Q-Sepharose chromatography allowed for the isolation of a homogeneous xylanase (PXII-1), which was 32.87 kDa according to MS analysis. The enzyme-specific activity towards soluble oat spelt xylan, which was found to be 490 IU/mg under optimum reaction conditions (50°C and pH 5.0-5.5), was 17-fold higher than that measured in the culture supernatant. Xylan reaction products were identified as xylobiose, xylotriose, and xylotetraose. K (m) values (mg ml(-1)) for soluble oat spelt and birchwood xylan were 11.8 and 9.45, respectively. Although PXII-1 showed 85% activity retention upon incubation at 50 °C and pH 5.0 for 20 days, incubation at pH 7.0 resulted in 50% activity loss within 3 days. PXII-1 stability at pH 7.0 was improved in the presence of 20 mM cysteine, which allowed for 85% activity retention for 25 days. This study on the production in high yields of a remarkably thermostable xylanase is of significance due to the central role that this class of biocatalyst shares, along with cellulases, for the much needed enzymatic hydrolysis of biomass. Furthermore, stable xylanases are important for the manufacture of paper, animal feed, and xylooligosaccharides.

show abstract

Section: Xylanase Hydrolytic Profilementioning

confidence: 99%

Purification and characterization studies of a thermostable β-xylanase from Aspergillus awamori

Teixeira

Siqueira

Souza

et al. 2010

J Ind Microbiol Biotechnol

View full text Add to dashboard Cite

show abstract

“…Some of these genes were then either overexpressed in Aspergillus species (13) or heterologously expressed in the yeast Saccharomyces cerevisiae and Pichia pastoris (20,14,19), but in most cases the low secretion levels did not allow complete purification and characterization of the enzymes. Nevertheless, heterologous expression of A. niger xylanase was of interest for the production of large quantities of a single xylanolytic enzyme.…”

Section: Hyperexpression Of Two Aspergillus Niger Xylanase Genementioning

confidence: 99%

“…niger reported (14,19). Characterization of the recombinant enzymes revealed the great difference between the recombinant XYNA1 and XYNB:…”

Section: Construction Of Expression Plasmid and Expression In Ecolimentioning

confidence: 99%

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

Yi¹,

Shi²,

Xu³

et al. 2010

Braz. J. Microbiol.

View full text Add to dashboard Cite

The analysis of individual gene product should enable to clarify the role of a particular enzyme in a complex xylanase system of A. niger. The two genes encoding precursors of co-produced endo-1,4--Dxylanases, xynA1 and xynB, were isolated from Aspergillus niger SCTCC 400264 (SCTCC, China) by using RT-PCR technique and then successfully expressed in Escherichia coli BL21. The nucleotide sequences of the xynA1 and xynB genes revealed that they were only 52.5% homology to each other.Characterization of the recombinant enzymes revealed the different properties: the specific activity of recombinant XYNA1 was 16.58 U/mg compared to 1201.7 U/mg for recombinant XYNB; The optimum temperature and pH of the recombinant XYNA1 were 35 ºC and 3.0, respectively, whereas the corresponding values for the recombinant XYNB were 55 ºC and 5.0, respectively; The recombinant XYNB showed much more thermostability than recombinant XYNA1; The recombinant XYNB showed 94% of maximal activity after incubating in water for 60 min at 60 ºC compared to no activity for recombinant XYNA1. Various metal ions had different effects on activity between the two recombinant xylanases.

show abstract

“…Even when A. fumigatus is commonly described as a pathogenic fungus [28], it is still used to produce enzymes such as xylanase [11], applied in industry [12]. Other possibility is to carry on studies about this interesting xylanase intending to insert and express the gene in non-pathogenic yeasts, as it was done by Liu et al [29]. …”

Section: Assays Of Cellulose Biobleaching Using Xylanasementioning

confidence: 99%

Production of xylanase by Aspergilli using alternative carbon sources: application of the crude extract on cellulose pulp biobleaching

Peixoto-Nogueira

Michelin

Betini

et al. 2008

J Ind Microbiol Biotechnol

View full text Add to dashboard Cite

The ability of xylanolytic enzymes produced by Aspergillus fumigatus RP04 and Aspergillus niveus RP05 to promote the biobleaching of cellulose pulp was investigated. Both fungi grew for 4-5 days in liquid medium at 40 degrees C, under static conditions. Xylanase production was tested using different carbon sources, including some types of xylans. A. fumigatus produced high levels of xylanase on agricultural residues (corncob or wheat bran), whereas A. niveus produced more xylanase on birchwood xylan. The optimum temperature of the xylanases from A. fumigatus and A. niveus was around 60-70 degrees C. The enzymes were stable for 30 min at 60 degrees C, maintaining 95-98% of the initial activity. After 1 h at this temperature, the xylanase from A. niveus still retained 85% of initial activity, while the xylanase from A. fumigatus was only 40% active. The pH optimum of the xylanases was acidic (4.5-5.5). The pH stability for the xylanase from A. fumigatus was higher at pH 6.0-8.0, while the enzyme from A. niveus was more stable at pH 4.5-6.5. Crude enzymatic extracts were used to clarify cellulose pulp and the best result was obtained with the A. niveus preparation, showing kappa efficiency around 39.6% as compared to only 11.7% for that of A. fumigatus.

show abstract

Expression of recombinant Aspergillus niger xylanase A in Pichia pastoris and its action on xylan

Cited by 41 publications

References 23 publications

Purification and characterization studies of a thermostable β-xylanase from Aspergillus awamori

Purification and characterization studies of a thermostable β-xylanase from Aspergillus awamori

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

Production of xylanase by Aspergilli using alternative carbon sources: application of the crude extract on cellulose pulp biobleaching

Contact Info

Product

Resources

About