Characterization of a novel amylosucrase gene from the metagenome of a thermal aquatic habitat, and its use in turanose production from sucrose biomass.

Abstract:

:Turanose is a natural isomer of sucrose. It is an emerging functional sweetener of the next generation. Turanose is catalytically synthesized from the sucrose biomass by employing amylosucrase enzyme. In this study, a novel gene encoding amylosucrase (Asmet) has been identified from the metagenome of a thermal aquatic habitat. Asmet exhibits 37-55% identity at the protein level with the known amylosucrases characterized till date. Asmet was cloned and expressed in Escherichia coli, followed by protein purification, and characterization. Asmet protein exhibited the maximum total activity at 9.0 pH and 60 °C temperature, whereas, 8.0 pH and 50 °C temperature were found optimum for transglycosylation activity. Asmet showed fairly high thermal tolerance at 50 °C. The conjugation of Asmet protein with functionalized iron nanoparticles significantly improved its thermal tolerance, showing hardly any loss in the enzyme's activity even after 72 h of heat (50 °C) exposure. The turanose yield of about 47% was achieved from 1.5 M sucrose, containing 0.5 M fructose in the reaction. Turanose was purified (˜95%) via a bio-physical process, and characterized by TLC, HPLC, and NMR. The novel amylosucrase gene was demonstrated to be a potential candidate for turanose production, utilizing various sucrose containing feedstocks.

journal_name

Enzyme Microb Technol

authors

Agarwal N,Narnoliya LK,Singh SP

doi

10.1016/j.enzmictec.2019.109372

subject

Has Abstract

pub_date

2019-12-01 00:00:00

pages

109372

eissn

0141-0229

issn

1879-0909

pii

S0141-0229(19)30109-7

journal_volume

131

pub_type

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