A Low-Temperature-Active Alkaline Pectate Lyase from Xanthomonas campestris ACCC 10048 with High Activity over a Wide pH Range
| Autor: | Kun Meng, Peilong Yang, Yaru Wang, Pengjun Shi, Huiying Luo, Tu Tao, Huoqing Huang, Peng Yuan, Bin Yao |
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| Rok vydání: | 2012 |
| Předmět: |
Intrinsic viscosity
Bioengineering Biology Molecular cloning Xanthomonas campestris medicine.disease_cause Applied Microbiology and Biotechnology Biochemistry Substrate Specificity Bacterial Proteins Affinity chromatography Enzyme Stability medicine Pectinase Molecular Biology Escherichia coli Polysaccharide-Lyases Temperature General Medicine Hydrogen-Ion Concentration biology.organism_classification Cold Temperature Kinetics Electrophoresis Pectate lyase Biotechnology Nuclear chemistry |
| Zdroj: | Applied Biochemistry and Biotechnology. 168:1489-1500 |
| ISSN: | 1559-0291 0273-2289 |
| Popis: | Alkaline pectate lyases are favorable for the textile industry. Here, we report the gene cloning and expression of a low-temperature-active alkaline pectate lyase (PL D) from Xanthomonas campestris ACCC 10048. Deduced PL D consists of a putative 27-residue signal peptide and a catalytic domain of 320 residues belonging to family PF09492. Recombinant PL D (r-PL D) produced in Escherichia coli was purified to electrophoretic homogeneity with a single step of Ni2+–NTA affinity chromatography and showed an apparent molecular weight of ~38 kDa. The pH and temperature optima of r-PL D were found to be 9.0 °C and 30 °C, respectively. Compared with its microbial counterparts, r-PL D had higher activity over a wide pH range (>45 % of the maximum activity at pH 3.0–12.0) and at lower temperatures (>35 % of activity even at 0 °C). The K m and V max values of r-PL D for polygalacturonic acid were 4.9 g l−1 and 30.1 μmol min−1 mg−1, respectively. Compared with the commercial compound pectinase from Novozymes, r-PL D showed similar efficacy in reducing the intrinsic viscosity of polygalacturonic acid (35.1 % vs. 36.5 %) and in bioscouring of jute (10.25 % vs. 10.82 %). Thus, r-PL D is a valuable additive candidate for the textile industry. |
| Databáze: | OpenAIRE |
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