A novel secreted metzincin metalloproteinase from Bacillus intermedius

The mprBi gene from Bacillus intermedius 3-19 encoding a novel secreted metalloproteinase was identified. The mpriBi gene was expressed in an extracellular proteinase-deficient Bacillus subtilis BG 2036 strain and the corresponding protein was characterized biochemically. The 19. kDa MprBi protein was purified to homogeneity and sequenced by mass spectroscopy and Edman degradation methods. Amino acid sequence analysis of MprBi identified an active site motif HEYGHNFGLPHD and a conserved structural component Met-turn, both of which are unique features of the metzincin clan. Furthermore, MprBi harbors a number of distinct sequence elements characteristic of proteinase domains in eukaryotic adamalysins. We conclude that MprBi and similar proteins from other Bacillus species form a novel group of metzincin metalloproteinases in prokaryotes. © 2010 Federation of European Biochemical Societies

The expression of the serine proteinase gene of Bacillus intermedius in Bacillus subtilis

The gene encoding for Bacillus intermedius serine proteinase was cloned and the complete nucleotide sequence was determined. Gene expression was explored in the protease-deficient strain Bacillus subtilis AJ73 during different stages of growth. Catabolite repression involved in control of proteinase expression during transition state and onset of sporulation was not efficient at the late stationary phase. Salt stress leads to induction of serine proteinase production during B. subtilis AJ73(pCS9) post-exponential growth. Expression of proteinase in B. subtilis deg-mutants may be controlled by DegU regulator. B. subtilis spo0-mutants failed to accomplish B. intermedius proteinase production. These data suggest complex network regulation of B. intermedius serine proteinase expression, including the action of spo0, degU, catabolite repression and demonstrate changes in control of enzyme biosynthesis at different stages of growth. © 2006 Elsevier GmbH. All rights reserved

Получение рекомбинантных нейротрофинов человека для биомедицинских исследований

The genes of nerve growth factor (NGF), brain neurotrophic factor (BDNF) and human neurotrophic factor 3 (NT-3) were cloned and expressed in Escherichia coli. Methods of purification and renaturation of the proneurotrophins were developed. It was shown that the recombinant pro-NGF, pro-BDNF and pro-NT-3 induce a differentiation of chicken dorsal root ganglia cell culture.Клонированы гены фактора роста нервов (NGF), нейротрофического фактора головного мозга (BDNF) и нейротрофического фактора 3 (NT-3) человека и осуществлена их экспрессия в клетках Escherichia coli. Разработаны схемы очистки и ренатурации соответствующих пробелков. Продемонстрировано, что рекомбинантные про-NGF, про-BDNF, про-NT-3 оказывают дифференцирующее действие на культуру спинномозговых ганглиев эмбрионов кур

Protealysin is not Secreted Constitutively

© 2019 Bentham Science Publishers. Background: Protealysin, a zinc metalloprotease of Serratia proteamaculans, is the prototype of a new group within the peptidase family M4. Protealysin-like proteases (PLPs) are widely spread in bacteria but are also found in fungi and archaea. The biological functions of PLPs have not been well studied, but published data showed the involvement of enzymes of this group in the interaction of bacteria with higher organisms, and most likely in the pathogenesis. Such functionality requires the release of the proteases from bacterial cells; however, the data on the cellular localization of PLPs are contradictory and no direct data of this kind have been published. Objective: Here, the protealysin cellular localization was studied for the first time using immunochemical methods. Methods and Results: We have produced polyclonal rabbit antibodies against the protealysin precursor. The enzyme was evaluated in cells and medium of periodic culture of S. proteamaculans 94 using Western blotting as well as the enzyme localization was analysed by immunoelectron microscopy. It was shown that more than 99% of the enzyme is in a cell-associated form. Protealysin is accumulated in cells as an inactive precursor. It matures only after the release from cells (after their lysis). Immunoelectron microscopy analysis of bacterial cells has revealed no specific localization of protealysin; it was evenly distributed in the cytoplasm. Conclusion: The data obtained suggest that S. proteamaculans protealysin and supposedly other protealysin-like proteases are not secreted constitutively and their release from bacteria is likely induced by a certain stimulus such as a contact with a eukaryotic cell. This finding is critical for further studies of the involvement of these enzymes in pathogenesis

Production of human recombinant neurotrophins for biomedical studies

The genes of nerve growth factor (NGF), brain neurotrophic factor (BDNF) and human neurotrophic factor 3 (NT-3) were cloned and expressed in Escherichia coli. Methods of purification and renaturation of the proneurotrophins were developed. It was shown that the recombinant pro-NGF, pro-BDNF and pro-NT-3 induce a differentiation of chicken dorsal root ganglia cell culture

Hugh Speer on Farm

Trustee Hugh Speer on his far

The expression of Bacillus intermedius glutamyl endopeptidase gene in Bacillus subtilis recombinant strains

The gene encoding for B. intermedius glutamyl endopeptidase (gseBi) has previously been cloned and its nucleotide sequence analyzed. In this study, the expression of this gene was explored in protease-deficient strain B. subtilis AJ73 during stationary phase of bacterial growth. We found that catabolite repression usually involved in control of endopeptidase expression during vegetative growth was not efficient at the late stationary phase. Testing of B. intermedius glutamyl endopeptidase gene expression with B. subtilis spo0-mutants revealed slight effect of these mutations on endopeptidase expression. Activity of glutamyl endopeptidase was partly left in B. subtilis ger-mutants. Probably, gseBi expression was not connected with sporulation. This enzyme might be involved in outgrowth of the spore, when germinating endospore converts into the vegetative cell. These data suggest complex regulation of B. intermedius glutamyl endopeptidase gene expression with contribution of several regulatory systems and demonstrate changes in control of enzyme biosynthesis at different stages of growth. © 2007 Springer Science+Business Media, Inc

Managing health outcomes through local governance

Has the devolution of responsibilities and finances to local governments in India produced positive impacts on health status and incomes of men and women? Using a national data set with details on health, health seeking behavior and local governance, we estimate a system of simultaneous equations to analyze the incidence of illness via a health production function and show that health care expenditures at the household level, choice of health care provider by members, and the incidence of illness will jointly impact individual incomes. The incidence of illness is significantly reduced by better access to drinking water, clean surroundings and awareness about health campaigns. Discussions of health issues in the Gram Sabha reduce illnesses for both men and women and their private health expenditures. Reserving the position of the village head to women leads to greater participation in Gram Sabha meetings; better problem resolution in water supply, sanitation and health; greater village health expenditures; and greater satisfaction with access to health. The increased expenditures of Panchayats on health care reduce the incidence of illness three times more for women than for men, but reduce their private health care expenditures about equally. It also shifts the choice of health care providers from private to public facilities, more so for women than for men. Family inheritance increase the use of private health care for both genders, while a woman’s individual land inheritance increases her use of both public and private health care. While women do not have lower access to health care or are discriminated within the household in terms of access to different providers, their earnings are adversely affected to a greater degree by illness compared to that of men. Women’s private health expenditures tend to improve their incomes more compared to that of men