Large-Scale Production of Glaciozyma antarctica Antifreeze Protein 1 (Afp1) by Fed-Batch Fermentation of Pichia pastoris

Mahzan Md Tab, Noor Haza Fazlin Hashim, Nazalan Najimudin, Nor Muhammad Mahadi, Farah Diba Abu Bakar, Abdul Munir Abd. Murad

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Antifreeze proteins (AFPs) are unique proteins that have an affinity toward ice. Due to their potential applications in the food and medical industries, many attempts have been made to produce AFPs in large quantities. In this study, recombinant Afp1 from Glaciozyma antarctica, a psychrophilic yeast, was overexpressed in a methylotrophic yeast, Pichia pastoris, followed by 5-L fermenter expression. The highest yield of Afp obtained from the fermentation was 39.5 mg/L when the cells were cultivated at 16∘C and at pH 5.0. Afp1 was produced as a glycoprotein (∼ 55 kDa) based on gel staining using a glycoprotein kit. Antifreeze activities of the recombinant Afp1 were exhibited through thermal hysteresis (TH) and recrystallization inhibition (RI) where the highest TH value recorded was at ∼0.5∘C at 10 mg/mL. This value is higher when compared to the recombinant Afp1 produced in Escherichia coli (0.08∘C) as well as the native antifreeze protein from G. antarctica (0.1∘C). Both TH and RI activities increased when higher protein concentrations were used. Effects of temperature on stability showed that Afp1 had lost its activity after being incubated at a temperature higher than 20∘C. The cryoprotective effects of Afp1 on cellulases showed that the treated cellulase retained up to ∼ 20% of its activity following several cycles of freeze–thawing. This indicates that Afp1 might act as a cryoprotective agent and has the potential for use in biotechnology applications.

Original languageEnglish
Pages (from-to)133-141
Number of pages9
JournalArabian Journal for Science and Engineering
Volume43
Issue number1
DOIs
Publication statusPublished - 1 Jan 2018

Fingerprint

batch fermentation
Pichia pastoris
Antarctica
hysteresis
recombinant proteins
heat
glycoproteins
antifreeze proteins
yeasts
cellulases
fermenters
endo-1,4-beta-glucanase
biotechnology
temperature
ice
proteins
fermentation
gels
Escherichia coli
industry

Keywords

  • Bioreactor
  • Cryopreservation
  • Heterologous expression
  • Ice-binding protein
  • Psychrophilic yeast
  • Thermal hysteresis

ASJC Scopus subject areas

  • General

Cite this

Large-Scale Production of Glaciozyma antarctica Antifreeze Protein 1 (Afp1) by Fed-Batch Fermentation of Pichia pastoris. / Tab, Mahzan Md; Hashim, Noor Haza Fazlin; Najimudin, Nazalan; Mahadi, Nor Muhammad; Abu Bakar, Farah Diba; Abd. Murad, Abdul Munir.

In: Arabian Journal for Science and Engineering, Vol. 43, No. 1, 01.01.2018, p. 133-141.

Research output: Contribution to journalArticle

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