Hybrid 2D/3D-quantitative structure-activity relationship studies on the bioactivities and molecular mechanism of antibacterial peptides.

Antimicrobial peptide (AMP) is the polypeptide, which protects the organism avoiding attack from pathogenic bacteria. Studies have shown that there were some antimicrobial peptides with molecular action mechanism involved in crossing the cell membrane without inducing severe membrane collapse, then interacting with cytoplasmic target-nucleic acid, and exerting antibacterial activity by interfacing the transmission of genetic information of pathogenic microorganisms. However, the relationship between the antibacterial activities and peptide structures was still unclear. Therefore, in the present work, a series of AMPs with a sequence of 20 amino acids was extracted from DBAASP database, then, quantitative structure-activity relationship (QSAR) methods were conducted on these peptides. In addition, novel antimicrobial peptides with  stronger antimicrobial activities were designed according to the information originated from the constructed models. Hence, the outcome of this study would lay a solid foundation for the in-silico design and exploration of novel antibacterial peptides with improved activity activities.

Antifungal mechanism of rose, mustard, and their blended essential oils against Cladosporium allicinum isolated from Xinjiang naan and its storage application.

AIMS: To reveal the inhibition mechanism of rose, mustard, and blended essential oils against Cladosporium allicinum isolated from Xinjiang naan, and investigate the effect of the three essential oils on oxidative damage and energy metabolism. METHODS AND RESULTS: Rose and mustard essential oils significantly inhibited mycelial growth and spore viability in a dose-dependent relationship. After essential oil treatment, the cell membrane permeability was altered, and significant leakage of intracellular proteins and nucleic acids occurred. SEM observations further confirmed the disruption of cell structure. ROS, MDA, and SOD measurements indicated that essential oil treatment induced a redox imbalance in C. allicinum, leading to cell death. As for energy metabolism, essential oil treatment significantly reduced Na+K+-ATPase, Ca2+Mg2+-ATPase, MDH activity, and CA content, impairing metabolic functions. Finally, storage experiments showed that all three essential oils ensured better preservation of naan, with mustard essential oil having the best antifungal effect. CONCLUSIONS: Rose and mustard essential oils and their blends can inhibit C. allicinum at multiple targets and pathways, destroying cell morphological structure and disrupting metabolic processes.

Changes in the structure and hydration properties of high-temperature peanut protein induced by cold plasma oxidation.

To improve the hydration properties of high-temperature pressed peanut protein isolate (HPPI), we investigated the effect of cold plasma (CP) oxidation on functional and structural properties. Compared to HPPI, the hydrated molecules number and the surface contact angle were significantly decreased at 70 W, from 77.2 × 109 to 17.7 × 109 and from 85.74° to 57.81°, respectively. The reduction of the sulfhydryl content and the increase of the disulfide bond and di-tyrosine content indicated that the structural transformation was affected by the oxidation effect. In terms of structural changes, a stretched tertiary structure, ordered secondary structure, and rough apparent structure were observed after CP treatment. Additionally, the enhancement of surface free energy and group content such as -COOH, -CO and -OH on the surface of HPPI contributed to the formation of hydrated crystal structures. In general, the oxidation effect of CP effectively improved the hydration properties of HPPI and broaden its application field.

Increased yield of 2-O-α-d-glucopyranosyl-l-ascorbic acid synthesis by α-glucosidase using rational design that regulating the ground state of enzyme and substrate complex.

BACKGROUND: α-Glucosidase (AG) is a bifunctional enzyme, it has a capacity to synthesize 2-O-α-d-glucopyranosyl-l-ascorbic acid (AA-2G) from l-ascorbic acid (L-AA) and low-cost maltose under mild conditions, but it can also hydrolyze AA-2G, which leads to low synthesis efficiency of AA-2G. MAIN METHODS AND MAJOR RESULTS: This study introduces a rational molecular design strategy to regulate enzymatic reactions based on inhibiting the formation of ground state of enzyme-substrate complex. Y215 was analyzed as the key amino acid site affecting the affinity of AG to AA-2G and L-AA. For the purpose of reducing the hydrolysis efficiency of AA-2G, the mutant Y215W was obtained by analyzing the molecular docking binding energy and hydrogen bond formation between AG and the substrates. Compared with the wild-type, isothermal titration calorimetry (ITC) results showed that the equilibrium dissociation constant (KD ) of the mutant for AA-2G was doubled; the Michaelis constant (Km ) for AA-2G was reduced by 1.15 times; and the yield of synthetic AA-2G was increased by 39%. CONCLUSIONS AND IMPLICATIONS: Our work also provides a new reference strategy for the molecular modification of multifunctional enzymes and other enzymes in cascade reactions system.

Characterization and correlation of dominant bacteria and volatile compounds in post-fermentation process of Ba-bao Douchi.

Ba-bao Douchi, traditionally produced and spontaneously fermented for 1-2 years, has a unique flavor. However, little information is known about microorganisms and volatile flavors, particularly their relationship. In this study, the aroma profiles including the key aroma compounds, and bacterial communities were characterized and the correlations between dominant bacterial genera with key aroma compounds were studied during the post-fermentation of Ba-bao Douchi. The research showed that 12 bacterial genera were identified as the dominant genus by high-throughput sequencing. A total of 84 volatile compounds were detected by HS-GC-IMS and HS-SPME-GC-MS. Odor activity value (OAV) and gas chromatography-Mass spectrometry-olfactometry (GC-MS-O) were combined to determine the key volatile compounds, and the main volatile compounds including ethyl hexanoate, ethyl heptanoate, isovaleraldehyde, (+)-α-pinene, beta-phellandrene, were found to be responsible for the strong fruitiness, chocolate fragrance, fresh scent flavor, and ginger flavor of Ba-bao Douchi. Pearson correlation analysis showed that 5 dominant bacterial genera were positively associated with > 6 key volatile compounds (p < 0.01, |r| > 0.7). Thus, these bacterial genera might have an effect on the biosynthesis of volatile components. This study provides a theoretical reference for revealing the functional microorganisms and improving the flavor quality of Ba-bao Douchi.

Rational molecular design for improving digestive enzyme resistance of beta-glucosidase from Trichoderma viride based on inhibition of bound state formation.

Beta-glucosidase (BGL1) is widely used in animal feed industries. However, degradation caused by digestive enzymes in the intestine hampers its application. Improving the resistance of feed enzymes against proteases is crucial in livestock farming. To improve the resistance of beta-glucosidase against pepsin and trypsin, a rational molecular design based on the inhibition of bound-state formation and secondary design was developed. The strategy includes: (1) prediction of the interaction surface of the pepsin-BGL1 complex structure, (2) prediction of key amino acids affecting the formation of the complex, (3) optimization of pepsin-resistant mutants by structural evaluation, (4) secondary molecular design based on pepsin-resistant mutants, and optimization of pepsin and trypsin-resistant mutants. Two BGL1 protein mutants (BGL1Q627C and BGL1Q627C/R543H/R646W) were constructed, and then mutated and wild-type BGL1s were expressed in Pichia pastoris. The half-life of BGL1Q627C and BGL1Q627C/R543H/R646W were 1.36 and 1.51 times that of the wild type upon pepsin exposure, respectively. For trypsin resistance, the half-life were 0.93 and 1.53 times that of the wild type, respectively. Compare to those of the wild type, most of the basic enzymatic properties of both mutants were not significantly changed except for increased Michaelis constants. The rational design method can be used as a guide for modifying other feed enzymes.

Lactic Acid Bacteria With Antioxidant Activities Alleviating Oxidized Oil Induced Hepatic Injury in Mice.

In order to screening new Lactic acid bacteria (LAB) strains to alleviating liver injury induced by oxidized oil, we isolated and screened LAB from Chinese fermented foods. Lactobacillus plantarum AR113, Pediococcus pentosaceus AR243, and Lactobacillus plantarum AR501 showed higher scavenging activity of α, α-Diphenyl-ß-Picrylhydrazyl (DPPH) free radical and hydrogen radical, stronger inhibition of lipid peroxidation, and better protective effect on yeast cells in vitro. In vivo, oral administration of L. plantarum AR501 improved the antioxidant status of injury mice induced by oxidized oil including decreasing lipid peroxidation, recovering activities of antioxidant enzymes. Meanwhile, the gene expression of Nuclear factor erythroid 2-related factor 2 (Nrf2) of L. plantarum AR501 group was markedly elevated, and several antioxidant genes such as glutathione S-transferase (GSTO1), heme oxygenase-1 (HO-1), Glutamate cysteine ligase (GCL), and NAD(P)H:quinone oxidoreductase-l (NQO1) were subsequently up regulated in mice liver. Therefore, L. plantarum AR501 could be considered as potential candidates for production of functional foods that can alleviate the oxidative damage induced by oxidized oil.

Lactobacillus plantarum AR501 Alleviates the Oxidative Stress of D-Galactose-Induced Aging Mice Liver by Upregulation of Nrf2-Mediated Antioxidant Enzyme Expression.

Lactic acid bacteria (LAB) have been used as ingredients of functional foods to promote health and prevent diseases because of their beneficial effects. This study aimed to investigate the antioxidative effects of LAB on the hepatotoxicity in D-galactose-induced aging mice. LAB were isolated from the traditional Chinese fermented foods and screened by the tolerance of hydrogen peroxide (H2 O2 ). Male ICR (Institute of Cancer Research) mice were subcutaneously injected with D-galactose for 5 weeks and then gastric gavage with LAB for 6 weeks. The results showed that Lactobacillus plantarum AR113 and AR501, and Pediococcu pentosaceus AR243 could tolerate up to 1.5 mM H2 O2 in vitro, and they could live through simulated gastrointestinal tract (GIT) to colonizing the GIT of host. In vivo, oral administration of L. plantarum AR113 and AR501 improved the antioxidant status of D-galactose-induced oxidative stress mice such as alleviated liver damages and reduced abnormal activities of superoxide dismutase, glutathione peroxidase, and catalase to normal levels. In addition, L. plantarum AR501 markedly elevated the gene expression of nuclear factor erythroid-2-related factor 2 and upregulated the expressions of several antioxidant genes such as glutathione reductase, glutathione S-transferase, glutamate-cysteine ligase catalytic subunit, glutamate-cysteine ligase modifier subunit, and NAD(P)H quinone oxidoreductase 1 in the liver of an aging mice. Therefore, L. plantarum AR501 could be a good candidate for producing antiaging functional foods.

Improvement of flavor profiles in Chinese rice wine by creating fermenting yeast with superior ethanol tolerance and fermentation activity.

Producing alcoholic beverages with novel flavor are desirable for winemakers. We created fermenting yeast with superior ethanol tolerance and fermentation activity to improve the flavor profiles of Chinese rice wine. Strategies of ethanol domestication, ultraviolet mutagenesis (UV) and protoplast fusion were conducted to create yeast hybrids with excellent oenological characteristic. The obtained diploid hybrid F23 showed a cell viability of 6.2% under 25% ethanol, whereas its diploid parental strains could not survive under 20% ethanol. During Chinese rice wine-making, compared to diploid parents, F23 produced 7.07%-12.44% higher yield of ethanol. Flavor analysis indicated that the total content of flavor compounds in F23 wine was 19.99-26.55% higher than that of parent wines. Specifically, F23 exhibited higher capacity in producing 2-phenylethanol, short-chain and long-chain fatty-acid ethyl-ester than diploid parents. Compared to diploid parents, F23 introduced more flavor contributors with odor activity values (OAVs) above one to Chinese rice wine, and those contributors were found with higher OAVs. Based on principal component analysis (PCA), the flavor characteristic of F23 wine was similar to each of parent wine. Additionally, sensory evaluation showed that F23 wine was highly assessed for its intensive levels in fruit-aroma, alcohol-aroma and mouthfeel. Hybrid F23 not only displayed superior flavor production and oenological performance in making Chinese rice wine, but also could act as potential "mixed-like" starter to enrich wine style and differentiation.