Characterization of a New Bifunctional and Cold-Adapted Polysaccharide Lyase (PL) Family 7 Alginate Lyase from Flavobacterium sp.

Abstract

Alginate oligosaccharides produced by enzymatic degradation show versatile physiological functions and biological activities. In this study, a new alginate lyase encoding gene alyS02 from Flavobacterium sp. S02 was recombinantly expressed at a high level in Yarrowia lipolytica, with the highest extracellular activity in the supernatant reaching 36.8 ± 2.1 U/mL. AlyS02 was classified in the polysaccharide lyase (PL) family 7. The optimal reaction temperature and pH of this enzyme were 30 °C and 7.6, respectively, indicating that AlyS02 is a cold-adapted enzyme. Interestingly, AlyS02 contained more than 90% enzyme activity at 25 °C, higher than other cold-adapted enzymes. Moreover, AlyS02 is a bifunctional alginate lyase that degrades both polyG and polyM, producing di- and trisaccharides from alginate. These findings suggest that AlyS02 would be a potent tool for the industrial applications.

Expression and Characterization of a Novel Cold-Adapted Chitosanase from Marine Renibacterium sp. Suitable for Chitooligosaccharides Preparation

“…To clarify the evolutionary relationship among CsnY and other chitosanases from bacteria, the phylogenetic tree construction was executed by the neighbor-joining method with MEGA 6.0 software, according to the protein sequences of related chitosanases obtained from National Center of Biotechnology Information (NCBI), Bethesda, MD, USA (https://www.ncbi.nlm.nih.gov/) [37]. The multiple sequences alignment was created by means of DNAMAN software (Lynnon Biosoft, Foster City, CA, USA) [38].…”

“…To further investigate the DPs of the oligosaccharides in the final product, the product solution was mixed with methanol (1:1, v/v), then quantitatively injected into an ESI-MS instrument (Bruker Esquire HCT, Billerica, MA, USA). The hydrolytic products were profiled in positive-ion mode under the following conditions: calibration dynamics, 2; cone voltage, 20.00 V; capillary voltage, 4.00 kV; desolvation temperature, 350 • C; source temperature, 150 • C; desolvation gas flow, 500 L/h; cone gas flow rate, 50 L/h; scan range, 100-1500 m/z [38].…”

Characterization of a New Biofunctional, Exolytic Alginate Lyase from Tamlana sp. s12 with High Catalytic Activity and Cold-Adapted Features

“…S02 and Alyw201 from Vibrio sp. W2 [ 19 , 20 ]. Moreover, the apparent Tm of rAlys1 determined by differential scanning calorimetry (DSC) was 56.0 °C ( Figure 4 c), which was consistent with the thermal stability of the rAlys1.…”

“…Notably, the enzyme activity of rAlys1 was only weakly affected by changing NaCl concentrations from 0.01 to 0.3 M (data not shown). The rAlys1 activity was increased with Na + , Mg 2+ , and K + , like alginate lyases from the PL7 family, which are essential for the enzyme activation for most alginate lyases [ 19 ].…”

Structure Characteristics, Biochemical Properties, and Pharmaceutical Applications of Alginate Lyases

“…Possessing low optimal temperature and low thermostability, cold-adapted alginate lyases catalyze the enzymatic reactions at low temperature, which can save energy and reduce the risk of microbial contaminations [70,78]. Moreover, such catalytic condition is crucial to improve the sustainability of enzyme utilization [79]. Therefore, making full use of cold-adapted alginate lyases can reduce the cost of industrial production to a certain extent.…”