Effects of cholesterol-lowering probiotic fermentation on the active components and in vitro hypolipidemic activity of sea buckthorn juice.

Sea buckthorn has lipid-lowering properties and is widely used in the development of functional foods. In this study, a probiotic (Lactobacillus plantarum, Lp10211) with cholesterol-lowering potential and acid and bile salt resistant was screened for the fermentation of sea buckthorn juice. Changes in the active ingredients, such as sugars and phenolics, before and after fermentation, as well as their in vitro lipid-lowering activities, were compared. The contents of reducing and total sugars decreased substantially after fermentation. Lp10211 primarily utilized fructose for growth and reproduction, with a utilization rate of 76.9%. The phenolic compound content of sea buckthorn juice increased by 37.06% after fermentation and protected the phenolic components from degradation (protocatechuic and p-coumaric acids) and produced new polyphenol (shikimic acid). Enhanced inhibition of pancreatic lipase activity (95.42%) and cholesterol micellar solubility (59.15%) was evident. The antioxidant properties of the fermentation broth were improved. Notably, Lp10211 preserved the color and reversed browning in sea buckthorn juice. The collective findings indicate that fermentation of sea buckthorn juice by Lp10211 may enhance the functional components and lipid-lowering activity of sea buckthorn, which may provide a new approach for the development of lipid-lowering foods.

Optimization of Microwave-Assisted Extraction Process of Total Flavonoids from Salicornia bigelovii Torr. and Its Hepatoprotective Effect on Alcoholic Liver Injury Mice.

The objective of this study was to determine the optimal extraction conditions for total flavonoids from S. bigelovii using microwave-assisted extraction and to analyze the protective effect of total flavonoids from S. bigelovii on alcoholic liver injury in mice. The optimization of the process conditions for the microwave-assisted extraction of total flavonoids from S. bigelovii was performed using response surface methodology, and an alcohol-induced acute liver injury model in mice was used to investigate the effects of different doses of total flavonoids (100 mg/kg, 200 mg/kg, and 400 mg/kg) on the levels and activities of serum alanine aminotransferase kits (ALT), glutamic oxaloacetic transaminase kits (AST), superoxide dismutase kits (SOD), glutathione peroxidase kits (GSH-Px), and malondialdehyde (MDA). We performed hematoxylin-eosin (H&E) staining analysis on pathological sections of mouse liver tissue, and qRT-PCR technology was used to detect the expression levels of the inflammatory factors IL-1 ß, IL-6, and TNF-α. The results revealed that the optimal extraction process conditions for total flavonoids in S. bigelovii were a material-to-liquid ratio of 1:30 (g/mL), an ethanol concentration of 60%, an extraction temperature of 50 °C, an ultrasound power of 250 W, and a yield of 5.71 ± 0.28 mg/g. Previous studies have demonstrated that the flavonoids of S. bigelovii can significantly inhibit the levels of ALT and AST in the serum (p < 0.001), reduce MDA levels (p < 0.001), increase the activity of the antioxidant enzymes SOD and GSH-Px (p < 0.001), and inhibit the IL-1 ß, IL-6, and TNF-α gene expression levels (p < 0.001) of inflammatory factors. The total flavonoids of S. bigelovii exert a protective effect against alcoholic liver injury by reducing the levels of inflammation, oxidative stress, and lipid peroxidation caused by alcohol. The results of this study lay the foundation for the high-value utilization of S. bigelovii and provide new resources for the development of liver-protective drugs.

A green method for decolorization of polysaccharides from alfalfa by S-8 macroporous resin and their characterization and antioxidant activity.

In this study, the decolorization conditions of polysaccharides extracted from alfalfa by S-8 macroporous adsorption resin were optimized through the response surface method, and the physicochemical properties and antioxidant activity of decolorized polysaccharides were investigated. The optimal decolorization conditions were determined to be as follows: the amount of S-8 macroporous adsorption resin was 1.4 g, the adsorption time was 2 h, and the adsorption temperature was 58 °C. Under these optimal conditions, a decolorization ratio of 71.43 ± 0.23% was achieved, which was consistent with the model hypothesis. The adsorption curve showed that S-8 macroporous adsorption resin adsorption of pigment molecules in alfalfa polysaccharides (APS) agreed with the Freundlich and pseudo-second-order equations, and the adsorption was a spontaneous endothermic process. High-performance liquid chromatography (HPLC) analysis of monosaccharide composition showed that APS was composed of mannose, glucose, galactose, arabinose and glucuronic acid in a molar ratio of 1.18 : 8.04 : 1.22 : 0.92 : 1. The results of antioxidant activity studies showed that APS had strong scavenging activity against ABTS, DPPH and hydroxyl radicals. This study will help to further understand the adsorption mechanism of macroporous resin on polysaccharide pigment molecules and lay a basis for evaluating their physiological activity.

Efficient whole cell biotransformation of tyrosol from L-tyrosine by engineered Escherichia coli.

An engineered Escherichia coli was constructed by co-expressing L-amino acid deaminase, α-keto acid decarboxylase, alcohol dehydrogenase, and glucose dehydrogenase through two plasmids for tyrosol production. The activity of the rate-limiting enzyme L-amino acid deaminase from Cosenzaea myxofaciens (CmAAD) toward tyrosine was improved by structure-guided modification. The enzyme activity of triple mutant CmAAD V438G/K147V/R151E toward tyrosine was ~5.12-fold higher than that of the wild-type CmAAD. Secondly, the plasmid copy numbers and the gene orders were optimized to improve the titer of tyrosol. Finally, the recombinant strain CS-6 transformed 10 mM tyrosine into 9.56 ± 0.64 mM tyrosol at 45 â„ƒ, and the space-time yield reached 0.478 mM·L-1·h-1. This study proposes a novel idea for the efficient and natural production of tyrosol, which has great potential for industrial application.

Heterologous overexpression of a novel halohydrin dehalogenase from Pseudomonas pohangensis and modification of its enantioselectivity by semi-rational protein engineering.

A gene encoding a halohydrin dehalogenase from Pseudomonas pohangensis (PpHHDH) was identified, synthesized and expressed in Escherichia coli. Subsequently, we used protein engineering to enhance the enzyme's enantioselectivity. We created two enantiocomplementary HHDH mutants, N160L and Q159L, which exhibited higher S- and R-selectivity toward PGE, respectively. The exchange of Leu at 159 for Gln led to a 2.3-fold increase in enantioselectivity (E-value of 22.2) compared to the wild-type. In addition, the N160L mutant displayed an inverted enantioselectivity (from ER = 9.8 to ES = 21.6) toward PGE. The wild-type PpHHDH and its variants were purified and characterized. They all displayed maximum activity at pH 7.5. The optimum temperature of mutant Q159L and N160L was increased from 35 °C to 40 °C. The wild-type PpHHDH and N160L mutant had good pH stability at pH 5.0-7.5, and Q159L showed an even wider range of pH tolerance, from pH 4.5 to pH 8.0. The mutants N160L and Q159L showed slightly better thermostability than wild-type PpHHDH. For most tested substrates, the two variants showed higher enantioselectivity. These findings further confirmed the importance of amino acid residues at positions 159 and 160 for the enantioselectivity of PpHHDH.