Effects of Bacillus velezensis zk1 on the physiology and metabolism of peaches

Bacillus velezensis zk1 is the dominant bacterium that causing rot in peaches. However, the mechanisms through that this bacterium causes rot have not been elucidated. Here, we explored the mechanisms of peach decay caused by B. velezensis zk1. The invasion of B. velezensis zk1 in peaches resulted in an increase in glucose and arabinose contents in fruit tissues. Moreover, the relative conductivity of the fruit reached 84% after 4 days of culture with bacterial invasion. With the destruction of cells, the malondialdehyde content increased, whereas the vitamin C, dialdehyde, flavonoid and total phenol contents decreased. Polyphenol oxidase, superoxide dismutase, catalase, peroxidase, ascorbate peroxidase and ammonia lyase activities also decreased. Overall, these findings demonstrated that B. velezensis zk1 infection damaged peach chloroplasts, mitochondria, respiratory chain activity and related free radical scavenging enzyme systems, thereby disrupting the normal physiological metabolism of peaches and causing rot.     

Plant Growth Promotion Function of Bacillus sp. Strains Isolated from Salt-Pan Rhizosphere and Their Biocontrol Potential against Macrophomina phaseolina

In recent decades, intensive crop management has involved excessive use of pesticides or fertilizers, compromising environmental integrity and public health. Accordingly, there has been worldwide pressure to find an eco-friendly and safe strategy to ensure agricultural productivity. Among alternative approaches, Plant Growth-Promoting (PGP) rhizobacteria are receiving increasing attention as suitable biocontrol agents against agricultural pests. In the present study, 22 spore-forming bacteria were selected among a salt-pan rhizobacteria collection for their PGP traits and their antagonistic activity against the plant pathogen fungus Macrophomina phaseolina. Based on the higher antifungal activity, strain RHFS10, identified as Bacillus vallismortis, was further examined and cell-free supernatant assays, column purification, and tandem mass spectrometry were employed to purify and preliminarily identify the antifungal metabolites. Interestingly, the minimum inhibitory concentration assessed for the fractions active against M. phaseolina was 10 times lower and more stable than the one estimated for the commercial fungicide pentachloronitrobenzene. These results suggest the use of B. vallismortis strain RHFS10 as a potential plant growth-promoting rhizobacteria as an alternative to chemical pesticides to efficiently control the phytopathogenic fungus M. phaseolina.     

直投式纳豆菌发酵剂制备工艺优化

为探索出一种工艺简便、成本低且活菌数高的直投式纳豆菌（Bacillus natto）发酵剂的制备方法，该研究以大米粉为基质替代传统的种子液培养基培养纳豆菌后直接将其烘干制成直投式纳豆菌发酵剂，通过单因素及正交试验优化直投式纳豆菌发酵剂的制备工艺。结果表明，用含1.5%蛋白胨的大米粉作为培养基，在接种量4.0%、加水量40%、培养温度43 ℃条件下培养纳豆菌28 h，菌体生长良好，将培养物烘干后制成的直投式纳豆菌发酵剂中的活菌数含量达4.71×109 CFU/g。此技术操作简便，可推广至工业生产中，简化生产过程。     

芽孢杆菌Bacillus sp.dwc-2对模拟地下水中U(Ⅵ)的还原行为研究

在厌氧条件下研究了西南地区一种典型土壤微生物芽孢杆菌Bacillus sp.dwc-2对模拟地下水中U(Ⅵ)的还原行为，重点考察了时间、无机阴离子、腐殖酸（HA）及富里酸（FA）对还原的影响，并利用TEM、EDS、SAED和XPS对还原后的样品进行了表征。结果表明：在pH=7.0、c_NaHCO3=5 mmol/L和T=303 K条件下，Bacillus sp.dwc-2对U(Ⅵ)的还原率随时间的增加而增加，24 h内最大还原率为12.2%，此后则随时间的增加逐渐降低；HA和FA对U(Ⅵ)的微生物还原行为有一定影响，其中HA和FA浓度为25 mg/L时，U(Ⅵ)的还原在24 h最明显，其还原率分别为14.2%和16.2%，但随着HA和FA浓度的继续增加，因在U(Ⅵ)离子与HA、FA形成的配合物表面形成致密的腐殖层，抑制了电子的转移，阻止了U(Ⅵ)的还原。此外，研究表明HCO₃^-也会抑制U(Ⅵ)的还原。TEM-SAED和XPS分析证实了还原过程中U(Ⅳ)的存在。上述结果可为真实环境中微生物还原U(Ⅵ)提供基础数据和参考。     

Safe preparation of beefy meaty peptide with Bacillus subtilis

Preparation of flavour peptides with microorganisms is an attractive choice for its large-scale production. However, beefy meaty peptide (BMP), as a research hotspot in the field of umami peptide, has been studied in few microorganisms, and furthermore, the safe preparation of BMP has not yet been reported. In this study, multi-copy BMP (8BMP) was successfully expressed and produced in the ‘generally recognized as safe’ Bacillus subtilis (B. subtilis). Firstly, 8BMP with the potentially intense umami was screened out based on molecular docking analysis. Then, it was successfully expressed in the B. subtilis 168 and verified through sensory evaluation. Finally, the scaling capacity of 8BMP was evaluated in a 5-L fermenter, with the highest yield (0.84 g L⁻¹) 6.4 times than that of shake flask, which is conductive for industrial production of BMP and other food peptides.     

Preparation of grape seed polypeptide and its calcium chelate with determination of calcium bioaccessibility and structural characterisation

Grape seed polypeptide (GSPP) was obtained by liquid fermentation with Bacillus subtilis TK15.015 from waste grape seed pomace. Its chelation ability with calcium ions was excellent. Simulation of in vitro gastrointestinal digestion showed that GSPP and calcium chelate (GSPP-Ca) slowly released calcium and decreased stomach stimulation. The solubility and transmission rate of GSPP-Ca after digestion in the gastrointestinal tract for 8 h maintained high levels of 74.4% and 46.4%, respectively, which far exceeded the levels achieved by other calcium supplements. Structural characterisation showed that calcium ions mainly react with binding sites present in peptides such as carboxyl oxygen, hydroxyl oxygen, amino nitrogen and amide bonds. GSPP-Ca may be a state in which a calcium-encapsulating complex formed, leaving calcium ions in a soluble state, suggesting its possible use as a high-quality calcium supplement.     

Gelatinised and hydrolysed corn starch is a cost-effective carbon source with higher production of L-lactic acid by Bacillus coagulans compared with glucose

L-lactic acid is an important organic acid widely used in pharmaceutical, food and textile industries. Bacillus coagulans BCS13002 can efficiently produce L-lactic acid with two kinds of carbon sources. BCS13002 produced L-lactic acid at a content of 10.23 ± 0.16 g/L and 11.67 ± 0.22 g/L, when glucose and gelatinised and hydrolysed corn starch （GHCS） were used, respectively. GHCS exhibits several advantages, including high yield of L-lactic acid and low cost. Proteomics analyses identified several key enzymes, which contributed to the higher production of L-lactic acid when GHCS was used as the carbon source. Those key enzymes were involved in the two-component system (SpoOF), pantothenate and CoA biosynthesis (pantothenate synthetase, 1.584-fold; dihydroxy-acid dehydratase, 1.517-fold), beta-alanine metabolism (1.605-fold) and valine, leucine and isoleucine biosynthesis (1.517-fold) pathways. This study provides a biological basis for using GHCS as a substitute of glucose in the production of L-lactic acid.     

粮油食品防霉微生物筛选及其活性物质初步研究

为开发新型防霉剂，降低粮油食品储藏、运输、流通等过程中霉变对其品质和食用安全的影响，研究筛选获得具有强防霉活性的微生物。采用抑菌圈法，以禾谷镰刀菌（Fusarium graminearum）为指示菌筛选具有防霉活性的菌株。利用形态学观察、生化特征鉴定和16S rDNA基因序列比对进行菌株鉴定；通过不同温度、pH和蛋白酶K处理对防霉活性物质进行初步分析。结果显示：通过初筛获得33株对禾谷镰刀菌具有强抑制活性的菌株，复筛后选择的7株菌归属为芽孢杆菌属（Bacillus）；活性物质分析发现7株菌的活性物质均不耐高温和强酸碱，同时蛋白酶K处理对其防霉效果没有明显影响，初步推断为肽类；通过抑菌谱测定发现菌株ASAG 62对常见霉菌（产黄青霉Penicillium flavum、黑曲霉Aspergillus Niger、赭曲霉Aspergillus ochre、黄曲霉Aspergillus flavus、禾谷镰刀菌F. graminearum）均具有良好的抑制效果。