Biochemical and genetic characterization of β-1,3 glucanase from a deep subseafloor Laceyella putida

Abstract

A β-1,3-glucanase (LpGluA) of deep subseafloor Laceyella putida JAM FM3001 was purified to homogeneity from culture broth. The molecular mass of the enzyme was around 36 kDa as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). LpGluA hydrolyzed curdlan optimally at pH 4.2 and 80 °C. In spite of the high optimum temperature, LpGluA showed relatively low thermostability, which was stabilized by adding laminarin, xylan, colloidal chitin, pectin, and its related polysaccharides. The gene for LpGluA cloned by using degenerate primers was composed of 1236 bp encoding 411 amino acids. Production of both LpGluA and a chitinase (LpChiA; Shibasaki et al. Appl Microbiol Biotechnol 98, 7845-7853, 2014) was induced by adding N-acetylglucosamine (GluNAc) to a culture medium of strain JAM FM3001. Construction of expression vectors containing the gene for LpGluA and its flanking regions showed the existence of a putative repressor protein.

Novel Anti-Fungal d-Laminaripentaose-Releasing Endo-β-1,3-glucanase with a RICIN-like Domain from Cellulosimicrobium funkei HY-13

“…When the enzyme reactions were conducted in the presence of tryptophan (Trp) residue-specific modifiers including Hg 2+ (1 mM) and N-bromosuccinimide (5 mM), rGluY was considerably inactivated by the compounds (Figure 4). Previously, a similar observation was also made with a GH64 endo-β-1,3-glucanase from S. matensis DIC-108 [6] reacted with curdlan in the presence of Hg 2+ (>20 µM), although the biocatalytic activity of Laceyella putida GH16 endo-β-1,3-glucanase [27] was only moderately suppressed by the metal ion. Taken together, the findings were consistent with the fact that N-bromosuccinimide and Hg 2+ ions oxidize the indole ring of strictly conserved Trp residues in the active site of endotype GH enzymes, which essentially participate in enzyme-substrate interaction [28,29].…”

Isolation of β-1,3-Glucanase-Producing Microorganisms from Poria cocos Cultivation Soil via Molecular Biology

“…The glycoside hydrolase β-1,3-glucanase, extensively distributed among plants, fungi, and bacteria, acts on 1,3-β-glucosidic bonds of structural β-1,3-glucans to hydrolyze or transfer glycosides [ 1 , 2 ]. Based on the hydrolysis position, β-1,3-glucanases are divided into endo-type (E.C.…”

Expression and Characterization of Recombinant β-1,3-Glucanase of Burkholderia cepacia [BiogenCC E76] Expressed in Escherichia coli Expression Systems

“…Elisitor β-1,3-glukan dapat dihasilkan oleh aktivitas β-1,3-glukanase, baik yang berasal dari tanaman (Mishra et al 2012) maupun mikroba endofitik (Shoresh et al 2010;Chen et al 2016). Induksi elisitor β-1,3-glukan terbukti meningkatkan ketahanan tanaman terhadap virus, bakteri, dan jamur patogen, seperti pada tanaman Arabidopsis (Ménard et al 2004), padi (Inui et al 1997), alfalfa (Kobayashi et al 2016), anggur (Aziz et al 2003, dan kedelai (Fliegmann et al 2004). Tanaman kedelai yang diinduksi dengan elisitor β-1,3-glukan dari P. sojae f.sp.…”