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Two water-extractable polysaccharide fractions designated as CWP (7. 37 × 105 Da) and CWP-0.2 (1.58 × 104 Da) were isolated and purified from chickpea (Cicer arietinum L.) seeds. The chemical structure of the two polysaccharides was characterized by various methods. Monosaccharide composition and methylation analysis showed that CWP was composed of Man and Glc in a molar ratio of 44.6:55.4, and CWP-0.2 was composed of Rha, Ara, Man, Glc, and Gal in a molar ratio of 10.6:23.3:5.2:4.9:56. Further structural characterization indicated that the main chain connection of CWP was → (2-β-d-Fruf-1) n →, and the main chain connection of CWP-0.2 was explored as → 2,4)-α-l-Rhap-(1 → 3)-α-d-Galp-(1 → with the branched chain of → 2,4)-α-l-Rhap-(1 → o-4. Besides, both CWP and CWP-0.2 had antioxidant and immunoregulatory activity in vitro, through scavenging DPPH· and ABTS·+ as well as stimulating production of NO, IL-6, TNF-α and MCP-1 in RAW 264.7 macrophages. CWP-0.2 revealed significantly higher bioactivity than CWP.

Abstract Herbicides are important tools for weed control in modern agriculture. In the search for potential herbicidal natural products from fungal species, harzianum A and B were identified from the biofertilizer fungus, Trichoderma brevicompactum. In the phytotoxicity assays on the dicot species Brassica chinensis, harzianum A and B reduced both shoot and root lengths at low concentrations and inhibited the seed germination at 2 μg mL−1. In addition, harzianum A and B also exhibited phytotoxicity against monocots, Oryza sativa L. cv. Nipponbare and Echinochloa crusgalli L. Beauv.. Compared with a common herbicide, 2,4-dichlorophenoxyacetic acid, harzianum A and B performed similar activity in a pot assay, and were more effective in post-emergence than pre-emergence conditions. Harzianum A and B have potential as efficient herbicide for controlling important dicotyledon and monocotyledon weeds at low concentrations. They can be sprayed in liquid form in both pre- and post-emergence conditions. Our results confirmed the importance of these molecules for the development of new herbicides.

Dietary intervention with plant protein is one of the main methods that is used to lessen the symptoms of malnutrition. Supplementary soy protein to undernourished weaning rats for 6 weeks significantly increased their body weight gain. After the intervention, the level of total short-chain fatty acids (SCFAs) was restored to 1,512.7 μg/g, while the level was only 637.1 μg/g in the 7% protein group. The amino acids (valine, isoleucine, phenylalanine, and tryptophan) increased in the colon, and vitamin B6 metabolism was significantly influenced in undernourished rats. The tryptophan and glycine-serine-threonine pathways were elevated, leading to an increase in the level of tryptophan and 5-hydroxytryptophan (5-HTP) in the serum. In addition, the relative abundance of Lachnospiraceae_NK4A136_group and Lactobacillus increased, while Enterococcus and Streptococcus decreased compared to undernourished rats. Overall, soy protein improved the growth of rats with malnutrition in early life by regulating gut microbiota and metabolites in the colon and serum.

The starch digestion processing of whole grain foods is associated with its health benefits in improving insulin resistance. This study modified the digestibility of whole quinoa flour (WQ) via heat-moisture treatment (HMT), HMT combined with pullulanase (HMT+P), HMT combined with microwave (HMT+M), and HMT combined with citric acids (HMT+A), respectively. Results showed that all the treatments significantly increased (p < 0.05) the total dietary fiber (TDF) content, amylose content, and resistant starch (RS) content, however, significantly decreased (p < 0.05) the amylopectin content and rapidly digestible starch (RDS) content of WQ. HMT+P brought the highest TDF content (15.3%), amylose content (31.24%), and RS content (15.71%), and the lowest amylopecyin content (30.02%) and RDS content (23.65%). HMT+M brought the highest slowly digestible starch (SDS) content (25.09%). The estimated glycemic index (eGI) was respectively reduced from 74.36 to 70.59, 65.87, 69.79, and 69.12 by HMT, HMT+P, HMT+M, and HMT+A. Moreover, a significant and consistent reduction in the heat enthalpy (ΔH) of WQ was observed (p < 0.05), after four treatments. All these effects were caused by changes in the starch structure, as evidenced by the observed conjunction of protein and starch by a confocal laser scanning microscope (CLSM), the decrease in relative crystallinity, and transformation of starch crystal.

Coarse cereals are rich in flavonoids, which are bioactive substances with a wide range of functions. Biotransformation is considered an emerging approach to methylate flavonoids, displaying prominent regio- and stereoselectivity. In the current study, liquiritigenin, naringenin, and hesperidin flavonoids were biotransformed using O-methyltransferases that were heterologously expressed in Saccharomyces cerevisiae BJ5464-NpgA. Nuclear magnetic resonance (NMR) spectroscopy was used together with high-resolution mass spectroscopy analysis to determine the structures of the resulting methylated transformants, and their antimicrobial and antiproliferation activities were also characterized. Among the five methylated flavonoids obtained, 7-methoxy-liquiritigenin had the strongest inhibitory effect on Candida albicans SC5314 (C. albicans SC5314), Staphylococcus aureus ATCC6538 (S. aureus ATCC6538), and Escherichia coli ATCC25922 (E. coli ATCC25922), which increased 7.65-, 1.49-, and 0.54-fold in comparison to the values of their unmethylated counterparts at 200, 250, and 400 μM, respectively. The results suggest that 3′-methoxyhesperetin showed the best antiproliferative activity against MCF-7 cells with IC50 values of 10.45 ± 0.45 µM, which was an increase of more than 14.35-fold compared to that of hesperetin. These results indicate that methylation enhances the antimicrobial activities and antiproliferative effects of flavonoids. The current study provides an experimental basis for further research on flavonoids as well as flavonoid-containing crops in the development of antimicrobial and anti-breast cancer drugs in addition to supplementary and health foods. The biotransformation method is ideal, as it represents a means for the sustainable production of bioactive flavonoids.

The widely used herbicide glyphosate targets 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). Glyphosate acetyltransferase (GAT) effectively detoxifies glyphosate by N-acetylation. With the aim of identifying a new strategy for development of glyphosate-tolerant crops, the plant expression vector pG2-GAT harboring gat and G2-aroA (encoding EPSPS) has been transformed into tobacco (Nicotiana tabacum) to develop novel plants with higher tolerance to glyphosate. Results from Southern and Western blotting analyses indicated that the target genes were integrated into tobacco chromosomes and expressed effectively at the protein level. Glyphosate tolerance was compared among transgenic tobacco plants containing gat, G2-aroA, or both genes. Plants containing both gat and G2-aroA genes were the most glyphosate-tolerant. This study has shown that a combination of different strategies may result in higher tolerance in transgenic crops, providing a new approach for development of glyphosate-tolerant crops.

Glycosylation is an efficient strategy to modulate the solubility, stability, bioavailability and bioactivity of drug-like natural products. Biological methods, such as whole-cell biocatalyst, promise a simple but highly effective approach to glycosylate biologically active small molecules with remarkable regio- and stereo-selectivity. Herein, we use the entomopathogenic filamentous fungus Isaria fumosorosea ACCC 37814 to biotransform a panel of phenolic natural products, including flavonoids and anthraquinone, into their glycosides. Six new flavonoid (4-O-methyl)glucopyranosides are obtained and structurally characterized using high resolution mass and nuclear magnetic resonance spectroscopic techniques. These compounds further expand the structural diversity of flavonoid glycosides and may be used in biological study.

An endo-β-1,4-glucanase gene, cel7A, was cloned from the thermophilic cellulase-producing fungus Neosartorya fischeri P1 and expressed in Pichia pastoris. The 1,410-bp full-length gene encodes a polypeptide of 469 amino acids consisting of a putative signal peptide at residues 1-20, a catalytic domain of glycoside hydrolase family 7 (GH7), a short Thr/Ser-rich linker and a family 1 carbohydrate-binding module (CBM 1). The purified recombinant Cel7A had pH and temperature optima of pH 5.0 and 60°C, respectively, and showed broad pH adaptability (pH 3.0-6.0) and excellent stability at pH3.0-8.0 and 60°C. Belonging to the group of nonspecific endoglucanases, Cel7A exhibited the highest activity on barley β-glucan (2020 ± 9 U mg-1), moderate on lichenan and CMC-Na, and weak on laminarin, locust bean galactomannan, Avicel, and filter paper. Under simulated mashing conditions, addition of Cel7A (99 μg) reduced the mash viscosity by 9.1% and filtration time by 24.6%. These favorable enzymatic properties make Cel7A as a good candidate for applications in the brewing industry.

A two-intein purification system was developed for the affinity purification of GFPmut3*, a mutant of green fluorescent protein. The GFPmut3* was sandwiched between two self-cleaving inteins. This approach avoided the loss of the target protein which may result from in vivo cleavage of a single intein tag. The presence of N- and C-terminal chitin-binding domains allowed the affinity purification by a single-affinity chitin column. After the fusion protein was expressed and immobilized on the affinity column, self-cleavage of the inteins was sequentially induced to release the GFPmut3*. The yield was 2.41 mg from 1 l of bacterial culture. Assays revealed that the purity was up to 98% of the total protein. The fluorescence and circular dichroism spectrum of GFPmut3* demonstrated that the purified protein retains the correctly folded structure and function. [ABSTRACT FROM AUTHOR]

Glyphosate, a powerful nonselective herbicide, acts as an inhibitor of the activity of the enzyme 5-enoylpyruvylshikimate-3-phosphate synthase (EPSPS) encoded by the aroA gene involved in aromatic amino acid biosynthesis. An Escherichia coli mutant AKM4188 was constructed by insertion a kanamycin cassette within the aroA coding sequence. The mutant strain is an aromatic amino acids auxotroph and fails to grow on M9 minimal media due to the inactive aroA. A DNA metagenomic library was constructed with samples from a glyphosate-polluted area and was screened by using the mutant AKM4188 as recipient. Three plasmid clones, which restored growth to the aroA mutant in M9 minimal media supplemented with chloramphenicol, kanamycin, and 50 m M glyphosate, were obtained from the DNA metagenomic library. One of them, which conferred glyphosate tolerance up to 150 m M, was further characterized. The cloned fragment encoded a polypeptide, designated RD, sharing high similarity with other Class II EPSPS proteins. A His-tagged RD fusion protein was produced into E. coli to characterize the enzymatic properties of the RD EPSP protein. [ABSTRACT FROM AUTHOR]