Expression and Characterization of a Novel β-Porphyranase from Marine Bacterium Wenyingzhuangia fucanilytica: A Biotechnological Tool for Degrading Porphyran

Abstract

Porphyra is one of the most consumed types of red algae. Porphyran is the major polysaccharide extracted from Porphyra, and it is composed of alternating 4-linked α-l-galactopyranose-6-sulfate (L6S) and 3-linked β-d-galactopyranose (G) residues. β-Porphyranases are promising tools for degrading porphyran; however, few enzymes have been reported, and the biochemical properties of porphyranases are still unclear. Here, a novel GH16 β-porphyranase, designated as Por16A_Wf, was cloned from Wenyingzhuangia fucanilytica and expressed in Escherichia coli. Its biochemical properties and hydrolysis pattern were characterized. Por16A_Wf exhibited stable activity on a wide pH scale from 3.5 to 11.0. Glycomics analysis using LC-MS revealed that Por16A_Wf specifically hydrolyzed the glycosidic linkage of G-L6S, whereas it tolerated 3,6-anhydro-α-l-galactopyranose and methyl-d-galactose in −2 and +2 subsites, respectively. Por16A_Wf could be applied as a biotechnological tool for tailoring porphyran, which would serve in directional preparation of its disaccharide, producing products with various molecular weights and facilitating investigation of the structural heterogeneity of Porphyra polysaccharides.

Discovery and Characterization of an Endo-1,3-Fucanase From Marine Bacterium Wenyingzhuangia fucanilytica: A Novel Glycoside Hydrolase Family

“…FunA activity was not significantly influenced by NaCl ( Supplementary Figure S4E ). The activities of marine polysaccharide enzymes were widely reported to depend on the concentration of NaCl, including Cgi82A ( Shen et al, 2017 ) and Por16A_Wf ( Zhang Y. et al, 2019 ) from W. fucanilytica . The salt-independent property of FunA would be beneficial to its practical applications.…”

Characterization of a Novel Porphyranase Accommodating Methyl-galactoses at Its Subsites

“…13 Recently, a porphyranase Por16A_Wf from Wenyingzhuangia fucanilytica CZ1127 T was characterized by our lab. 14 It tolerates LA and GMe at subsites −2 and +2, respectively. On the basis of the amino acid sequence similarity, PorA, PorB, BpGH16B, and Por16A_Wf are classified into the glycoside hydrolase (GH) family 16, while BpGH86A belongs to the GH86 family.…”

Marine Polysaccharides: Occurrence, Enzymatic Degradation and Utilization

“…Later a new β‐porphyranase from Bacteroides plebeius from family GH86 was identified [60] . Similar studies indicated the importance of GH16 enzymes in the degradation of porphyran by the investigation of further GH16 porphyranases [85,89] . Nevertheless, these enzymes are not sufficient on their own for the complete degradation of porphyran (Figure 2).…”

“… [60] Similar studies indicated the importance of GH16 enzymes in the degradation of porphyran by the investigation of further GH16 porphyranases. [ 85 , 89 ] Nevertheless, these enzymes are not sufficient on their own for the complete degradation of porphyran (Figure 2 ). They require a synergistic cleavage of several side group‐removing enzymes which deprotect the polysaccharide chain from functional groups and thereby enable further degradation by the porphyranases.…”