Heterologous expression of Aspergillus aculeatus endo-polygalacturonase in Pichia pastoris by high cell density fermentation and its application in textile scouring

• Published in: BMC biotechnology Vol. 17; no. 1; p. 15
• Main Authors: Abdulrachman, Dede, Thongkred, Paweena, Kocharin, Kanokarn, Nakpathom, Monthon, Somboon, Buppha, Narumol, Nootsara, Champreda, Verawat, Eurwilaichitr, Lily, Suwanto, Antonius, Nimchua, Thidarat, Chantasingh, Duriya
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 16.02.2017; BioMed Central
• Subjects: alkali treatment; Analysis; Aspergillus - enzymology; Aspergillus - genetics; Aspergillus aculeatus; Batch Cell Culture Techniques - methods; batch fermentation; Bioreactors; Bioreactors - microbiology; bioscouring; Cellulose; Citrus; Cotton; cotton fabric; Cotton fabrics; Cotton Fiber; Detergents - chemistry; Detergents - metabolism; E coli; Energy consumption; Enzymatic analysis; Enzymes; Fermentation; Gene expression; Glycerol; heterologous gene expression; Komagataella pastoris; Materials Testing; pectins; Pichia - enzymology; Pichia - genetics; Pichia - growth & development; Plasmids; polygalacturonase; Polygalacturonase - biosynthesis; Polygalacturonase - chemistry; Polygalacturonase - genetics; Potassium; Protein Engineering - methods; Proteins; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; tensile strength; Textiles; wettability; Yeast; Selangor Malaysia; United States > US; Malaysia; Cotton fabric; Acidic pectinase; High cell density fermentation; Bioscouring; Heterologous expression
• ISSN: 1472-6750; 1472-6750
• DOI: 10.1186/s12896-017-0334-9

Removal of non-cellulosic impurities from cotton fabric, known as scouring, by conventional alkaline treatment causes environmental problems and reduces physical strength of fabrics. In this study, an endo-polygalacturonase (EndoPG) from Aspergillus aculeatus produced in Pichia pastoris was evaluated for its efficiency as a bioscouring agent while most current bioscouring process has been performed using crude pectinase preparation. The recombinant EndoPG exhibited a specific activity of 1892.08 U/mg on citrus pectin under the optimal condition at 50 °C, pH 5.0 with a V and K of 65,451.35 μmol/min/mL and 15.14 mg/mL, respectively. A maximal activity of 2408.70 ± 26.50 U/mL in the culture supernatant was obtained by high cell density batch fermentation, equivalent to a 4.8 times greater yield than that from shake-flask culture. The recombinant enzyme was shown to be suitable for application as a bioscouring agent, in which the wettability of cotton fabric was increased by treatment with enzyme at 300 U/mL scouring solution at 40 °C, pH 5.0 for 1 h. The bio-scoured fabric has comparable wettability to that obtained by conventional chemical scouring, but has higher tensile strength. The work has demonstrated for the first time functions of A. aculeatus EndoPG on bioscouring in eco-textile processing. EndoPG alone was shown to possess effective scouring activity. High expression level and homogeneity could be achieved in bench-scale bioreactor.