An α-1,6-and α-1,3-linked glucan produced by Leuconostoc citreum ABK-1 alternansucrase with nanoparticle and film-forming properties

Scientific Reports
Karan WangpaiboonRath Pichyangkura

Abstract

Alternansucrase catalyses the sequential transfer of glucose residues from sucrose onto another sucrose molecule to form a long chain polymer, known as "alternan". The alternansucrase-encoding gene from Leuconostoc citreum ABK-1 (Lcalt) was successfully cloned and expressed in Escherichia coli. Lcalt encoded LcALT of 2,057 amino acid residues; the enzyme possessed an optimum temperature and pH of 40 °C and 5.0, respectively, and its' activity was stimulated up to 2.4-fold by the presence of Mn2+. Kinetic studies of LcALT showed a high transglycosylation activity, with Km 32.2 ± 3.2 mM and kcat 290 ± 12 s-1. Alternan generated by LcALT (Lc-alternan) harbours partially alternating α-1,6 and α- 1,3 glycosidic linkages confirmed by NMR spectroscopy, methylation analysis, and partial hydrolysis of Lc-alternan products. In contrast to previously reported alternans, Lc-alternan can undergo self-assembly, forming nanoparticles with an average size of 90 nm in solution. At concentrations above 15% (w/v), Lc-alternan nanoparticles disassemble and form a high viscosity solution, while this polymer forms a transparent film once dried.

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Citations

Sep 30, 2020·World Journal of Microbiology & Biotechnology·Peteris ZikmanisPavels Semjonovs
May 16, 2020·The Journal of Biological Chemistry·Manon MolinaMagali Remaud-Siméon
Jan 7, 2021·Organic & Biomolecular Chemistry·Trichada RatthachagPanuwat Padungros
Apr 4, 2021·International Journal of Molecular Sciences·Karan WangpaiboonRath Pichyangkura
Aug 4, 2021·International Journal of Biological Macromolecules·Thanapon CharoenwongpaiboonRobert A Field
May 28, 2019·Journal of Agricultural and Food Chemistry·Franziska MünkelDaniel Wefers

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Methods Mentioned

BETA
PCR
NMR
Dynamic light scattering

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