Purification of β-mannanase derived from Bacillus subtilis ATCC 11774 using ionic liquid as adjuvant in aqueous two-phase system.

Autor: Aziz NFHA; Bioprocess Technology, School of Industrial Technology, Universiti Sains Malaysia, 11800 Penang, Malaysia., Abbasiliasi S; Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia., Ng HS; Faculty of Applied Sciences, UCSI University, 56000 Cheras, Kuala Lumpur, Malaysia., Phapugrangkul P; Thailand Institute of Scientific and Technological Research, Bio-Sciences Department, 35, Moo3, Klong 5, Klong Luang, Pathumthani 12120, Thailand., Bakar MHA; Bioprocess Technology, School of Industrial Technology, Universiti Sains Malaysia, 11800 Penang, Malaysia., Tam YJ; Laboratory of Immunotherapeutic and Vaccine Technology (LIVES), Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia., Tan JS; Bioprocess Technology, School of Industrial Technology, Universiti Sains Malaysia, 11800 Penang, Malaysia. Electronic address: jooshun@usm.my.
Jazyk: angličtina
Zdroj: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences [J Chromatogr B Analyt Technol Biomed Life Sci] 2017 Jun 15; Vol. 1055-1056, pp. 104-112. Date of Electronic Publication: 2017 Apr 17.
DOI: 10.1016/j.jchromb.2017.04.029
Abstrakt: The partitioning of β-mannanase derived from Bacillus subtilis ATCC 11774 in aqueous two-phase system (ATPS) was studied. The ATPS containing different molecular weight of polyethylene glycol (PEG) and types of salt were employed in this study. The PEG/salt composition for the partitioning of β-mannanase was optimized using response surface methodology. The study demonstrated that ATPS consists of 25% (w/w) of PEG 6000 and 12.52% (w/w) of potassium citrate is the optimum composition for the purification of β-mannanase with a purification fold (PF) of 2.28 and partition coefficient (K) of 1.14. The study on influences of pH and crude loading showed that ATPS with pH 8.0 and 1.5% (w/w) of crude loading gave highest PF of 3.1. To enhance the partitioning of β-mannanase, four ionic liquids namely 1-butyl-3-methylimidazolium tetrafluoroborate ([Bmim]BF 4 ), 1-ethyl-3-methylimidazolium tetrafluoroborate ([Emim]BF 4 ), 1-butyl-3-methylimidazolium bromide ([Bmim]Br), 1-ethyl-3-methylimidazolium bromide ([Emim]Br) was added into the system as an adjuvant. The highest recovery yield (89.65%) was obtained with addition of 3% (w/w) of [Bmim]BF 4 . The SDS-PAGE analysis revealed that the β-mannanase was successfully recovered in the top phase of ATPS with the molecular size of 36.7kDa. Therefore, ATPS demonstrated a simple and efficient approach for recovery and purification of β-mannanase from fermentation broth in one single-step strategy.
(Copyright © 2017 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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