D-Phenylglycine aminotransferase (D-PhgAT) from Pseudomonas stutzeri ST-201 is useful for enzymatic synthesis of enantiomerically pure D-phenylglycine. However, its low protein solubility prevents its application at high substrate concentration. With an aim to increase the protein solubility, the N-terminus of D-PhgAT was genetically fused with short peptides (A1 α- helix, A2 α-helix, and ALAL, which is a hybrid of A1 and A2) from a ferredoxin enzyme of a halophilic archaeon, Halobacterium salinarum. The fused enzymes A1-D-PhgAT, A2-D-PhgAT, and ALAL-D-PhgAT displayed a reduced pI and increased in solubility by 6.1-, 5.3-, and 8.1- fold in TEMP (pH 7.6) storage, respectively, and 5-, 4.5-, and 5.9-fold in CAPSO (pH 9.5) reaction buffers, respectively, compared with the wild-type enzyme (WT-D-PhgAT). In addition, all the fused D-PhgAT displayed higher enzymatic reaction rates than the WT-DPhgAT at all concentrations of L-glutamate monosodium salt used. The highest rate, 23.82 ± 1.47 mM/h, was that obtained from having ALAL-D-PhgAT reacted with 1,500 mM of the substrate. Moreover, the halophilic fusion significantly increased the tolerance of D-PhgAT in the presence of NaCl and KCl, being slightly in favor of KCl, where under the same condition at 3.5 M NaCl or KCl all halophilic-fused variants showed higher activity than WT-D-PhgAT.

Aim: Mutation occurs in the prostate cell genes, leading to abnormal prostate proliferation and ultimately cancer. Prostate cancer (PC) is one of the most common cancers amongst men, and its prevalence worldwide increases relative to men's age. About 16% of the world's cancers are the result of microbes in the human body.Impaired population balance of symbiosis microbes in the human reproductive system is linked to PC development.Discussion: With the advent of metagenomics science, the genome sequence of the microbiota of the human body has been unveiled. Therefore, it is now possible to identify a higher range of microbiome changes in PC tissue via the Next Generation Technique, which will have positive consequences in personalized medicine. In this review, we intend to question the role of metagenomics studies in the diagnosis and treatment of PC. Conclusion:The microbial imbalance in the men's genital tract might have an effect on prostate health. Based on next-generation sequencing-generated data, Proteobacteria, Firmicutes, Actinobacteria, and Bacteriodetes are the nine frequent phyla detected in a PC sample, which might be involved in inducing mutation in the prostate cells that cause cancer.

Evolution of life in extreme environments entails wide adaptations. Amongst these environments, those with high salt concentration are abodes to so-called halophilic microorganisms. The unique key properties of halophilic proteins not only enable them to survive in hostile and precluding environments, but also make their enzymes as precious biocatalysts in pharmaceutical, fine chemical processes, industries, food processing, detergents, waste treatments, etc., where harsh conditions prevent favorable reactions by normal enzymes. Unfortunately, due to lack of deep knowledge and proper equipment, up to know only a limited number of extremozymes have characterized and applied in appropriate industries. Discovery of new extremophiles or genetically engineering of mesophilic enzymes will open new horizons to novel applications of these invaluable enzymes in biotechnology and biotransformation.

Background D-Phenylglycine aminotransferase (D-PhgAT) is highly beneficial in pharmaceutical biotechnology. Like many other enzymes, D-PhgAT suffers from low stability under harsh processing conditions, poor solubility of substrate, products and occasional microbial contamination. Incorporation of miscible organic solvents into the enzyme’s reaction is considered as a solution for these problems; however, native D-PhgAT is not significantly stable in such solvents. Objective Halophiles are known to survive and withstand unsavory habitats owing to their proteome bios. In the current study, with an eye on further industrial applications, we examined the performance and thermostability of four halophilic peptides fused D-PhgAT variants in reaction mixtures of various proportions of different miscible organic solvents and various temperatures as well as desiccation. Materials and Methods Plasmid constructs from the previous study (Two alpha helixes and loops between them from Halobacterium salinarum ferredoxin enzyme fused at N-terminus domain of D-PhgAT) expressed in Escherichia coli and then D-PhgAT purified. Purified proteins were subjected to various proportions of miscible organic solvents, different temperatures, and desiccation and then performance and thermostability monitored. Results Study confirmed increased C 50 of all halophilic fused D-PhgAT variants, where the highest C 50 observed for ALAL-D-PhgAT (30.20±2.84 %V/V). Additionally, all halophilic fused variants showed higher thermostability than the wild-type D-PhgAT in the presence of different fractions of acetone, N,N -Dimethylformamide and isopropanol in aqueous binary media, while zero activity observed at the presence of methanol. Conclusion Our results suggest that applying this new technique could be invaluable for making enzymes durable in discordant industrial conditions.