Activity-Based Screening of Soil Samples from Nyingchi, Tibet, for Amylase-Producing Bacteria and Other Multifunctional Enzyme Capacities.

Despite the interest in Tibetan soil as a promising source of functional enzymes with potential biotechnological applications, few studies have considered the screening and identification of amylase producing bacteria from Tibetan soil. Amylase has many applications in the food and feed industries, textile and biofuel production, and biomedical engineering. The area of amylase with specific properties is attracting growing attention because of its better application to various industrial conditions. This study aims to screen and identify amylase-producing strains from soil samples collected in Nyingchi, Tibet, and then explore whether the bacterial isolates are superior for unique enzymes. In this paper, a total of 127 amylase producing bacteria were isolated by activity-based screening of six Tibetan soil samples. The 16S rRNA gene survey then identified four major phyla, namely, firmicutes, bacteroidetes, proteobacteria, and actinobacteria, which were differentiated into twelve genera with a dominance of Bacillus (67.72%), followed by Pseudomonas (8.66%). Microbial diversity analysis revealed that the amylase-producing bacterial community of the Kadinggou forest soil sample showed the best variety (the Simpson index was 0.69 and the Shannon index was 0.85). The amylase activity assay of the bacterial isolates showed a mean of 0.66 U/mL at 28°C and pH 5.2. Based on the effect of temperatures and pHs on amylase activity, several bacterial isolates can produce thermophilic (50°C), psychrophilic (10°C), acidophilic (pH 4.2), and alkaliphilic (pH 10.2) amylases. Furthermore, four bacterial isolates were screened for amylase, protease, and esterase activities, which indicated multifunctional enzyme capacities. The present study is expected to contribute to our understanding of Tibetan microbial resources and their potential for scientific research and industrial applications.

Gastrodin suppresses BACE1 expression under oxidative stress condition via inhibition of the PKR/eIF2α pathway in Alzheimer's disease.

The expression of ß-site APP-cleaving enzyme 1 (BACE1) is increased in the brain of late-onset sporadic Alzheimer's disease (AD) and oxidative stress may be the potential cause of this event. The phenolic glucoside gastrodin (Gas), a main component of a Chinese herbal medicine Gastrodia elata Blume, has been demonstrated to display antioxidant activity and suppresses BACE1 expression. However, the mechanisms by which Gas suppresses BACE1 expression are not clear. Morris water maze test was performed to assess the effect of Gas treatment on memory impairments in Tg2576 mice. The level of oxidative stress in the brain of Tg2576 mice was determined by measuring the superoxide dismutase (SOD) activity, catalase (CAT) activity, and the levels of malondialdehyde (MDA) and ROS. In vivo and in vitro, we detected the expression levels of BACE1, pPKRThr446, PKR, pPERKThr981, PERK, peIF2αSer51, and eIF2α using western blot analysis. We found that Gas improved learning and memory abilities of Tg2576 transgenic mice and attenuated intracellular oxidative stress in hippocampi of Tg2576 mice. We discovered that the expression levels of BACE1, activated PKR (pPKRThr446) and activated eIF2α (peIF2αSer51) were elevated in the brains of Tg2576 mice and hydrogen peroxide (H2O2)-stimulated SH-SY5Y cells. Moreover, peptide PKR inhibitor (PRI) and Gas down-regulated BACE1 expression in Tg2576 mice and H2O2-stimulated SH-SY5Y cells by inhibiting activation of PKR and eIF2α. Gas alleviates memory deficits in mice and suppresses BACE1 expression by inhibiting the protein kinase/Eukaryotic initiation factor-2α (PKR/eIF2α) pathway. The research suggested that Gas may develop as an drug candidate in neurodegenerative diseases.