Effects of mutation of Asn694 in Aspergillus niger α-glucosidase on hydrolysis and transglucosylation

Aspergillus niger α-glucosidase (ANG), a member of glycoside hydrolase family 31, catalyzes hydrolysis of α-glucosidic linkages at the non-reducing end. In the presence of high concentrations of maltose, the enzyme also catalyzes the formation of α-(1→6)-glucosyl products by transglucosylation and it is used for production of the industrially useful panose and isomaltooligosaccharides. The initial transglucosylation by wild-type ANG in the presence of 100 mM maltose [Glc(α1–4)Glc] yields both α-(1→6)- and α-(1→4)-glucosidic linkages, the latter constituting ~25% of the total transfer reaction product. The maltotriose [Glc(α1–4)Glc(α1–4)Glc], α-(1→4)-glucosyl product disappears quickly, whereas the α-(1→6)-glucosyl products panose [Glc(α1–6)Glc(α1–4)Glc], isomaltose [Glc(α1–6)Glc], and isomaltotriose [Glc(α1–6)Glc(α1–6)Glc] accumulate. To modify the transglucosylation properties of ANG, residue Asn694, which was predicted to be involved in formation of the plus subsites of ANG, was replaced with Ala, Leu, Phe, and Trp. Except for N694A, the mutations enhanced the initial velocity of the α-(1→4)-transfer reaction to produce maltotriose, which was then degraded at a rate similar to that by wild-type ANG. With increasing reaction time, N694F and N694W mutations led to the accumulation of larger amounts of isomaltose and isomaltotriose than achieved with the wild-type enzyme. In the final stage of the reaction, the major product was panose (N694A and N694L) or isomaltose (N694F and N694W).

     

Hydrothermal synthesis, characterization, and KOH activation of carbon spheres from glucose

Carbon spheres (CSs) with controllable sizes and rich in oxygen-containing groups were fabricated using a simple hydrothermal treatment of glucose. The effects of the hydrothermal parameters, including the concentration of glucose, reaction temperature, duration, and the second hydrothermal treatment were investigated. The obtained CSs were then activated using KOH for the eventual preparation of porous carbon spheres. A scanning electron microscope was used to characterize the morphology and size of the CSs. Fourier-transform infrared spectroscopy and X-ray photoelectron spectroscopy were used to analyze the functional surface groups. N₂ adsorption–desorption isotherms were used to analyze the porous structure of the CS. The results revealed that the morphologies and size distribution of the CSs can be controlled by adjusting the experimental parameters. A hydrothermal temperature between 180 and 190 °C over 4–5 h was suitable for CS formation. Under these conditions, the size of the CS increased with the concentration of glucose. Mono-dispersed CSs with good morphologies and large numbers of oxygen-containing functional groups (primarily –OH and CO) can be obtained using a 0.3 mol/L glucose solution that is hydrothermally treated at 190 °C for 4 h. The resulting CSs sizes were about 350 nm in diameter. After a second hydrothermal treatment, the sizes of CSs grew nearly 250 nm without damage to its morphology or broadening of their size distribution. Porous CSs with perfectly spherical shapes and fully developed structures (S_BET = 1282.8 m²/g, V_micro = 0.44 cm³/g) could then be obtained via KOH activation.     

Antrodiella pendulina and A. citripileata spp. nov. (Basidiomycota,Polyporales) from subtropical and tropical China

Antrodiella pendulina H. S. Yuan and A. citripileata H. S. Yuan spp. nov. from southern China are illustrated and described. Antrodiella pendulina has disciform basidiocarps, a trimitic hyphal system, cylindrical basidiospores and occur exclusively on angiosperm twigs. Antrodiella citripileata has effused‐reflexed to pileate and lemon‐colored basidiocarps, very small pores, thick‐walled skeletal hyphae with a wide lumen, and small ellipsoid basidiospores.     

High-throughput screening for Streptomyces antibiotic biosynthesis activators

A genomic cosmid library of Streptomyces clavuligerus was constructed and transferred efficiently by conjugation to Streptomyces lividans, and 12 distinct groups of overlapping cosmid clones that activated the silent actinorhodin biosynthesis gene cluster were identified. This generally applicable high-throughput screening procedure greatly facilitates the identification of antibiotic biosynthesis activators.