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

نویسندگان

  • Dede Abdulrachman
  • Paweena Thongkred
  • Kanokarn Kocharin
  • Monthon Nakpathom
  • Buppha Somboon
  • Nootsara Narumol
  • Verawat Champreda
  • Lily Eurwilaichitr
  • Antonius Suwanto
  • Thidarat Nimchua
  • Duriya Chantasingh
چکیده

BACKGROUND 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. RESULTS 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 max and K m 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. CONCLUSION 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.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Cloning and heterologous expression of Laccase in pichia pastoris and determination some of biochemical properties

Laccase (EC 1.10.3.2) are multi-copper oxidase which catalyze the oxidation aromatic and non- aromatic compounds with electron reduction of molecular oxygen to water. Nucleotide sequence of laccase (accession number : ) was optimized according codon preference of Pichia pastoris. Gene was synthesized and cloned into pPICZalpha A. laccase under control of AOX1 promoter was transformed to P.pasto...

متن کامل

High-Level Expression of Endo-β-N-Acetylglucosaminidase H from Streptomyces plicatus in Pichia pastoris and Its Application for the Deglycosylation of Glycoproteins

Endo-β-N-acetylglucosaminidase H (Endo H, EC3.2.1.96) is a glycohydrolase that is widely used in the study of glycoproteins. The present study aimed to assess the effect of high-level endo-β-N-acetylglucosaminidase H expression in Pichia pastoris. The DNA coding sequence of this enzyme was optimized based on the codon usage bias of Pichia pastoris and synthesized through overlapping PCR. This n...

متن کامل

Production of Recombinant Human Granulocyte-Colony Stimulating Factor by Pichia pastoris

Human granulocyte-colony stimulating factor (hG-CSF) cDNA was expressed in the methylotrophic yeast Pichia pastoris under the control of the alcohol oxidase (AOX1) promoter. An expression vector for hG-CSF secretion was constructed using vector pPIC9. Higher levels of hG-CSF was obtained using a P. pastoris Mut+ (methanol utilization fast) phenotype. The effects of environmental factors such as...

متن کامل

Comparison of biochemical properties of recombinant phytase expression in the favorable methylotrophic platforms of Pichia pastoris and Hansenula polymorpha

Phytic acid is the dominant form of phosphorous in plant seeds. However, phytic acid cannot beutilized by animals and causes them serious phosphate deficiency. Utilization of phytase as ananimal feed additive can affect liberation of phosphor and its mineral availability. In this study,heterologous expression of the A. niger phyA gene was investigated in the methylotrophic yeastsP. pastoris and...

متن کامل

Heterologous Expression of Bovine Prochymosin in Pichia pastoris GS115

Objectives: In present research we evaluate the expression of this critical enzyme in a eukaryotic system for future use in cheese industry. Materials and Methods: We have cloned bovine prochymosin gene in methylotrophic yeast, P. pastoris, using pPIC9K as an expression vector. The recombinant plasmid was transformed into the host by electroporation, and it was expre...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره 17  شماره 

صفحات  -

تاریخ انتشار 2017