A novel ß-1,3-glucanase Gns6 from rice possesses antifungal activity against Magnaporthe oryzae.

As members of the pathogenesis-related protein (PR)-2 family, ß-1,3-glucanases play pivotal roles in plant defense. Previous study showed that the rice genome contains 16 genes encoding putative ß-1,3-glucanases, and the ß-1,3-glucanases in subfamily A were deduced to be involved in plant defense. However, there was limited direct evidence. In this study, the expression of rice ß-1,3-glucanases Gns2-Gns6 belonging to subfamily A in rice plant infection with Magnaporthe oryzae was investigated, and the enhanced expression of Gns6 during infection confirmed its crucial role in the defense of rice seedlings. Enzymological characterization revealed that Gns6 preferentially hydrolyzed laminarin, pachymaran, and yeast glucan. The ß-1,3; 1,6-glucanase Gns6 exhibited a specific activity of 1.2 U/mg with laminarin as the substrate. In addition, Gns6 could hydrolyze laminarin via an endo-type mechanism, yielding a series of oligosaccharides with various degrees of polymerization that are known immune elicitors in plants. Moreover, Gns6 exhibited a significant inhibitory effect against the formation of the germ tubes and appressoria, with potential applications in plant protection. Taken together, this study shows that Gns6 is an essential effector in the defensive response of rice against pathogenic fungi.

Structural and digestion properties of potato starch modified using an efficient starch branching enzyme AqGBE.

Modified potato starch with slower digestion may aid the development of new starch derivatives with improved nutritional values, and strategies to increase nutritional fractions such as resistant starch (RS) are desired. In this study, a correspondence between starch structure and enzymatic resistance was provided based on the efficient branching enzyme AqGBE, and modified starches with different amylose content (Control, 100%; PS1, 90%; PS2, 72%; PS3, 32%; PS4, 18%) were prepared. Through SEM observation, NMR and X-ray diffraction analyses, we identified that an increased proportion of α-1,6-linked branches in potato starch changes its state of granule into large pieces with crystallinity. Molecular weight and chain-length distribution analysis showed a decrease of molecular weight (from 1.1 × 106 to 1.1 × 105 g/mol) without an obvious change of chain-length distribution in PS1, while PS2-4 exhibited an increased proportion of DP 6-12 with a stable molecular weight distribution, indicating a distinct model of structural modification by AqGBE. The enhancement of peak viscosity was related to increased hydrophobic interactions and pieces state of PS1, while the contents of SDS and RS in PS1 increased by 37.7 and 49.4%, respectively. Our result provides an alternative way to increase the RS content of potato starch by branching modification.

Characterization of an acidic pectin methylesterase from Paenibacillus xylanexedens and its application in fruit processing.

A pectinase-producing bacterial isolate, identified as Paenibacillus xylanexedens SZ 29, was screened by using the soil dilution plate with citrus pectin and congo red. A pectin methylesterase gene (Pxpme) was cloned and expressed in Escherichia coli. The gene coded for a protein with 334 amino acids and a calculated molecular mass of 36.76 kDa. PxPME showed the highest identity of 32.4% with the characterized carbohydrate esterase family 8 pectin methylesterase from Daucus carota. The recombined PxPME showed a specific activity with 39.38 U/mg against citrus pectin with >65% methylesterification. The optimal pH and temperature for PxPME activity were 5.0 and 45 °C. Its Km and Vmax value were determined to be 1.43 mg/mL and 71.5 µmol/mg·min, respectively. Moreover, PxPME could increase the firmness of pineapple cubes by 114% when combined with CaCl2. The acidic and mesophilic properties make PxPME a potential candidate for application in the fruit processing.