| RRC ID |
84534
|
| 著者 |
Maati J, Prazeres DM, Grąz M, Wiater A, Jarosz-Wilkołazka A, Smaali I.
|
| タイトル |
Heteroxylan hydrolysis by a recombinant cellulase-free GH10 xylanase from the alkaliphilic bacterium Halalkalibacterium halodurans C-125.
|
| ジャーナル |
Arch Microbiol
|
| Abstract |
The search for affordable enzymes with exceptional characteristics is fundamental to overcoming industrial and environmental constraints. In this study, a recombinant GH10 xylanase (Xyn10-HB) from the extremely alkaliphilic bacterium Halalkalibacterium halodurans C-125 cultivated at pH 10 was cloned and expressed in E. coli BL21(DE3). Removal of the signal peptide improved the expression, and an overall activity of 8 U/mL was obtained in the cell-free supernatant. The molecular weight of purified Xyn10-HB was estimated to be 42.6 kDa by SDS-PAGE. The enzyme was active across a wide pH range (5-10) with optimal activity recorded at pH 8.5 and 60 °C. It also presented good stability with a half-life of 3 h under these conditions. Substrate specificity studies showed that Xyn10-HB is a cellulase-free enzyme that conventionally hydrolyse birchwood and oat spelts xylans (Apparent Km of 0.46 mg/mL and 0.54 mg/mL, respectively). HPLC analysis showed that both xylans hydrolysis produced xylooligosaccharides (XOS) with a degree of polymerization (DP) ranging from 2 to 9. The conversion yield was 77% after 24 h with xylobiose and xylotriose as the main end-reaction products. When assayed on alkali-extracted wheat straw heteroxylan, the Xyn10-HB produced active XOS with antioxidant activity determined by the DPPH radical scavenging method (IC50 of 0.54 mg/mL after 4 h). Owing to its various characteristics, Xyn10-HB xylanase is a promising candidate for multiple biotechnological applications.
|
| 巻・号 |
206(6)
|
| ページ |
261
|
| 公開日 |
2024-5-16
|
| DOI |
10.1007/s00203-024-03982-w
|
| PII |
10.1007/s00203-024-03982-w
|
| PMID |
38753095
|
| MeSH |
Bacterial Proteins / chemistry
Bacterial Proteins / genetics
Bacterial Proteins / metabolism
Cloning, Molecular
Disaccharides
Endo-1,4-beta Xylanases* / chemistry
Endo-1,4-beta Xylanases* / genetics
Endo-1,4-beta Xylanases* / metabolism
Enzyme Stability
Escherichia coli / genetics
Escherichia coli / metabolism
Glucuronates / metabolism
Hydrogen-Ion Concentration
Hydrolysis
Kinetics
Molecular Weight
Oligosaccharides / metabolism
Recombinant Proteins* / chemistry
Recombinant Proteins* / genetics
Recombinant Proteins* / isolation & purification
Recombinant Proteins* / metabolism
Substrate Specificity
Xylans* / metabolism
|
| リソース情報 |
| 一般微生物 |
JCM9153 |
