Cell-envelope proteinases from lactic acid bacteria: Biochemical features and biotechnological applications

Proteins displayed on the cell surface of lactic acid bacteria (LAB) perform diverse and important biochemical roles. Among these, the cell-envelope proteinases (CEPs) are one of the most widely studied and most exploited for biotechnological applications. CEPs are important players in the proteolytic system of LAB, because they are required by LAB to degrade proteins in the growth media into peptides and/or amino acids required for the nitrogen nutrition of LAB. The most important area of application of CEPs is therefore in protein hydrolysis, especially in dairy products. Also, the physical location of CEPs (i.e., being cell-envelope anchored) allows for relatively easy downstream processing (e.g., extraction) of CEPs. This review describes the biochemical features and organization of CEPs and how this fits them for the purpose of protein hydrolysis. It begins with a focus on the genetic organization and expression of CEPs. The catalytic behavior and cleavage specificities of CEPs from various LAB are also discussed. Following this, the extraction and purification of most CEPs reported to date is described. The industrial applications of CEPs in food technology, health promotion, as well as in the growing area of water purification are discussed. Techniques for improving the production and catalytic efficiency of CEPs are also given an important place in this review.     

Conditions stimulating neutral detergent fiber degradation by dosing branched-chain volatile fatty acids. I: Comparison with branched-chain amino acids and forage source in ruminal batch cultures

Three experiments assessed branched-chain volatile fatty acid (BCVFA) stimulation of neutral detergent fiber (NDF) disappearance after 24 h of incubation in batch cultures derived from ruminal fluid inocula that were enriched with particulate-phase bacteria. In experiment 1, a control was compared with 3 treatments with isomolar doses of all 3 BCVFA (plus valerate), all 3 branched-chain AA (BCAA), or half of each BCVFA and BCAA mix with either alfalfa or grass hays (50%) and ground corn grain (50%). A portion of the BCAA and BCVFA doses were enriched with ¹³C, and valerate (also enriched with ¹³C) was added with BCVFA. Although BCAA yielded a similar production of BCVFA compared with dosing BCVFA, equimolar substitution of BCVFA for BCAA decreased the percentage of N in bacterial pellets when alfalfa hay was fed but increased N when grass hay was fed. Substituting BCVFA for BCAA increased total fatty acid (FA) concentration with alfalfa hay. Dosing of BCAA or BCVFA did not affect total branched-chain FA, iso-FA, or anteiso-FA percentages in bacterial total FA, whereas numerous individual FA isomers and their ¹³C enrichments were affected by these treatments. Increasing recovery of the ¹³C dose from respective labeled BCVFA primers indicated facilitated BCVFA uptake and incorporation into FA compared with BCAA, whereas increased recovery of ¹³C from labeled BCAA in the bacteria pellet but not in the FA fraction suggested direct assimilation into bacterial protein. The BCVFA and valerate were dosed in varying combinations that either summed to 4 mM (experiment 2) or had only 1 mM no matter what combination (experiment 3). In general, grass hay was more responsive to stimulation in NDF digestibility by BCVFA than was alfalfa hay, which was attributed to the higher degradable protein in the latter. The net production of the BCVFA (after subtracting dose) was affected by source and combination of BCVFA. Isovalerate dosing tended to increase its own net production; in contrast, isobutyrate seemed to be used more when it was added alone, but 2-methylbutyrate seemed to be preferred over isobutyrate when 2-methylbutyrate was added. Results supported potential interactions, including potential feedback in production from feed BCAA or increased concentration-dependent competition for dosed BCVFA into cellular products. Under our conditions, the BCVFA appear to be more readily available than BCAA, probably because of regulated BCAA transport and metabolism. Valerate consistently provided no benefit. Using nonparametric ranking, all 3 BCVFA or either isovalerate or isobutyrate (both yielding iso-FA) should be combined with 2-methylbutyrate (yielding anteiso-FA) as a potential opportunity to improve NDF digestibility when rumen-degraded BCAA are limited in diets to decrease environmental impact from N in waste.     

Growth inhibition of Fusarium graminearum and deoxynivalenol detoxification by lactic acid bacteria and their application in sourdough bread

Eight lactic acid bacteria (LAB) strains were screened for their ability to inhibit Fusarium graminearum (F. graminearum) growth and remove deoxynivalenol (DON). Furthermore, the selected LAB strains were applied as starter cultures to evaluate their performance during the breadmaking process. Lactobacillus plantarum (L. plantarum) AR524 exhibited strong inhibitory activity against F. graminearum growth in a pH-dependent manner and removed up to 40.9% of DON, mainly through cell wall binding. Compared with the control, L. plantarum AR524 inhibited 60.19% of F. graminearum growth and removed 50.53% of DON during breadmaking. The texture properties of the bread were also remarkably improved by L. plantarum AR524, and its shelf life was extended from 3 days to 6 days without the application of any chemical preservatives. Therefore, L. plantarum AR524 shows excellent potential for application as a bioprotective starter culture in bakery products.     

发酵十字花科蔬菜中硫代葡萄糖苷代谢研究进展

硫代葡萄糖苷（简称硫苷）是十字花科蔬菜中一类重要的次生代谢物。在蔬菜发酵过程中，硫苷被代谢为异硫氰酸盐等活性产物，这些代谢产物具有抗氧化、抗癌、调节肠道菌群等多种生理功能。乳酸菌发酵对硫苷的代谢机制包括促进硫苷从蔬菜中释放，以及使其发生降解，且代谢作用既与乳酸菌种属相关，也与发酵体系的盐含量、金属离子、pH值以及蔬菜加热预处理相关。通过总结梳理发酵蔬菜硫苷代谢研究进展，并解析影响因素，为提高发酵蔬菜中活性硫苷代谢产物含量，提升发酵蔬菜的健康效应提供理论指引。     

The Peculiar Structure of Acetobacter pasteurianus CIP103108 LPS Core Oligosaccharide

Acetobacter pasteurianus, a member of the Alphaproteobacteria, is an acetic acid-producing bacterium present on sugar-rich substrates such as such as fruits, flowers and vegetables and traditionally used in the production of fermented food. The preferred living habitat associated with acid conditions makes the structure of the bacterial cell wall interesting to study, due to expected uncommon features. We have used a combination of chemical, analytical and NMR spectroscopy approaches to define the complete structure of the core oligosaccharide from A. pasteurianus CIP103108 LPS. Interestingly, the core oligosaccharide displays a high concentration of negatively charged groups, structural features that might contribute to reinforcing the bacterial membrane.     

Effect of Cultivation Parameters on Fermentation and Hydrogen Production in the Phylum Thermotogae

The phylum Thermotogae is composed of a single class (Thermotogae), 4 orders (Thermotogales, Kosmotogales, Petrotogales, Mesoaciditogales), 5 families (Thermatogaceae, Fervidobacteriaceae, Kosmotogaceae, Petrotogaceae, Mesoaciditogaceae), and 13 genera. They have been isolated from extremely hot environments whose characteristics are reflected in the metabolic and phenotypic properties of the Thermotogae species. The metabolic versatility of Thermotogae members leads to a pool of high value-added products with application potentials in many industry fields. The low risk of contamination associated with their extreme culture conditions has made most species of the phylum attractive candidates in biotechnological processes. Almost all members of the phylum, especially those in the order Thermotogales, can produce bio-hydrogen from a variety of simple and complex sugars with yields close to the theoretical Thauer limit of 4 mol H₂/mol consumed glucose. Acetate, lactate, and L-alanine are the major organic end products. Thermotagae fermentation processes are influenced by various factors, such as hydrogen partial pressure, agitation, gas sparging, culture/headspace ratio, inoculum, pH, temperature, nitrogen sources, sulfur sources, inorganic compounds, metal ions, etc. Optimization of these parameters will help to fully unleash the biotechnological potentials of Thermotogae and promote their applications in industry. This article gives an overview of how these operational parameters could impact Thermotogae fermentation in terms of sugar consumption, hydrogen yields, and organic acids production.     

Effects of extracellular proteases and its inhibitors on the gel characteristics of soy protein induced by lactic acid bacteria

The purpose of this study was to analyse the relationship between extracellular protease activity and gel characteristics of soybean protein gel induced by lactic acid bacteria (LAB), the influence of protease inhibition on the gel characteristics was discussed through SDS-PAGE, rheological properties and microstructure. The results showed that the pH value of Lactobacillus casei decreased slowly, but the gel could be formed at 2 h, maybe its higher extracellular protease activity improved the gel characteristics. The addition of ethylenediaminetetraacetic acid (10 mmol L⁻¹) reduced the gel quality, when the protease activity was inhibited, the gel quality would decrease, even if there were macromolecular crosslinkers in the system. This study showed that extracellular protease had an important influence on gel characteristics and provided a theoretical basis for the application of protease producing LAB in fermentation-induced soybean gel foods.     

Hsp100 Molecular Chaperone ClpB and Its Role in Virulence of Bacterial Pathogens

This review focuses on the molecular chaperone ClpB that belongs to the Hsp100/Clp subfamily of the AAA+ ATPases and its biological function in selected bacterial pathogens, causing a variety of human infectious diseases, including zoonoses. It has been established that ClpB disaggregates and reactivates aggregated cellular proteins. It has been postulated that ClpB’s protein disaggregation activity supports the survival of pathogenic bacteria under host-induced stresses (e.g., high temperature and oxidative stress), which allows them to rapidly adapt to the human host and establish infection. Interestingly, ClpB may also perform other functions in pathogenic bacteria, which are required for their virulence. Since ClpB is not found in human cells, this chaperone emerges as an attractive target for novel antimicrobial therapies in combating bacterial infections.     

汉江上游干支流沉积物细菌群落多样性和影响因素

为了解汉江上游干支流沉积物细菌多样性以及确定性过程和随机性过程在沉积物细菌群落构建过程中的相对重要性,基于Illumina高通量测序技术,分析了环境因子对细菌群落组成的影响,采用非度量多维尺度(NMDS)排序探究了季节之间沉积物细菌群落的差异,并结合中性群落模型和标准化随机率量化了确定性过程和随机过程对群落构建的影响。结果表明:汉江上游及其支流细菌群落主要由变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、蓝藻门(Cyanophyta)、浮霉菌门(Planctomycetes)和酸杆菌门(Acidobacteria)等组成；细菌群落在不同季节有显著差异；地理距离和环境因子对细菌群落结构影响较小,确定性过程并未在细菌群落组成中起到主导作用；随机过程很大程度上影响了群落在秋季和春季的组成,是沉积物细菌群落构建的主导因素。