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Coconut water (Cocos nucifera L.) possesses natural hydrating qualities, functional health properties and various nutritional benefits. Lactobacillus species AAF-1 has been isolated and purified from coconut water. The isolated culture was screened for the production of extracellular secreted enzymes having industrial value. Growth of the Lactobacillus sp. AAF-1 on MRS agar containing respective substrates showed that the Lactobacillus sp. AAF-1 is the potential producer of amylase, protease, cellulase and beta galactosidase. Further cell free filtrates of the Lactobacillus sp. AAF-1 showed large zone of hydrolysis of starch, gelatin, cellulose and lactose that reflected the extracellular production of the above enzymes in culture medium at 50 °C. It has been observed that activities after 72 h of incubation in cell free filtrates of protease, amylase, cellulase and beta galactosidase as the zone of respective substrate hydrolysis measured as 10, 8, 6 and 10 mm respectively. Thus, Lactobacillus sp. AAF-1 could be the potential producer of the enzymes having industrial value. [ABSTRACT FROM AUTHOR]

Antibody‐mediated enzyme formation is a phenomenon first described in 1968 and further studied by molecular Immunologists and Biochemists over the following five decades. The present review is made mainly by analyzing the 27 articles concerned with AMEF that appeared over the course of 47 years, commenting 16 original figures selected to be re‐printed in AMEF's Legacy. We, the reviewers, started by revisiting our own "insider's" experience of discovery, and followed by considering all results, our own and of members of other AMEF Labs. We had planned to conclude the review by correlating the various AMEF mutants to a detailed knowledge of the consensus betaGal structure. However, we became aware of several "robust" papers, published between 1989 and 2014, by authors outside of AMEF Labs. We familiarly called this surge: "The Second Wave" and adorned it with a doodle in Hokusai style. We were thrilled and happy to take them on board and properly examined their data. A team of this second wave had imagined unique uses for AMEF, and new doors to modern biotechnology. Another one had used AMEF as Tool and Marker to attain high levels of crystallography, solving puzzles of conformation, and ultimate structure. Together, they doubled our motivation to review AMEF. Serendipity gives us back the pleasure of finding, a treat at any age. [ABSTRACT FROM AUTHOR]

β-Galactosidase was immobilized on modified nanosilver reduced graphene oxide (Ag@rGO) nanocomposite prepared by in vitro synthesis using same enzyme. The effectiveness factor, η value of the immobilized enzyme was calculated to be 0.968, suggesting enhancement in enzyme activity after immobilization. The morphological structure of the crosslinked biopolymer was analyzed using electron microscopy and other characterization techniques. The kinetics displayed a decrease in K m value from 0.50 to 0.44 mmol L−1 while there was an increase in V max values from 0.031 to 0.039 μmol min−1 mL−1. The immobilized enzyme retained 85% activity after its 10th repeated use. Inhibition constant (K i) value suggests galactose to be a more potent inhibitor of the enzyme. Despite the inhibitory potential of these hydrolysis products, the immobilized enzyme preparation retained 44.2% activity in the presence of both inhibitory sugars. The as-synthesized nanobiocatalyst was found quite effective in hydrolyzing 89% of lactose from whey. Hence, this nanobiocatalyst can be used in removing lactose from dairy waste, whey before releasing it into the water bodies. Also, the cytotoxicity and genotoxicity of Ag@rGO NC was assessed on human blood lymphocytes using flow cytometry and comet assay, respectively. [Display omitted] • Formulation of nanosilver reduced graphene oxide nanocomposite using chemical synthesis approach. • Successful immobilization of β galactosidase on in vitro synthesized support using physical adsorption • Improved resistance to inhibition by the hydrolysis products by the immobilized enzyme • Synthesized support was found to be non toxic and biocompatible. • Enhanced lactose hydrolyzing potential of the immobilized enzyme [ABSTRACT FROM AUTHOR]

Morquio B disease is an attenuated phenotype within the spectrum of beta galactosidase (GLB1) deficiencies. It is characterised by dysostosis multiplex, ligament laxity, mildly coarse facies and heart valve defects due to keratan sulphate accumulation, predominantly in the cartilage. Morquio B patients have normal neurological development, setting them apart from those with the more severe GM1 gangliosidosis. Morquio B disease, with an incidence of 1:250.000 to 1:1.000.000 live births, is very rare. Here we report the clinical-biochemical data of nine patients. High amounts of keratan sulfate were detected using LC-MS/MS in the patients' urinary samples, while electrophoresis, the standard procedure of qualitative glycosaminoglycans analysis, failed to identify this metabolite in any of the patients' samples. We performed molecular analyses at gene, gene expression and protein expression levels, for both isoforms of the GLB1 gene, lysosomal GLB1, and the cell-surface expressed Elastin Binding Protein. We characterised three novel GLB1 mutations [c.75 + 2 T > G, c.575A > G (p.Tyr192Cys) and c.2030 T > G (p.Val677Gly)] identified in three heterozygous patients. We also set up a copy number variation assay by quantitative PCR to evaluate the presence of deletions/ insertions in the GLB1 gene. We propose a diagnostic plan, setting out the specific clinical- biochemical and molecular features of Morquio B, in order to avoid misdiagnoses and improve patients' management. • Morquio B is a very rare disease and we here report nine affected patients. • Clinical- biochemical and molecular aspects of Morquio B condition are detailed. • LC/MS-MS is proven the gold standard for the detection of keratan sulfate in urines. • Both beta- galactosidase and elastin binding proteins were evaluated. • We present a general paper and define an accurate diagnostic plan for Morquio B. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Ganoderma lucidum (Lingzhi or Reishi) is a medicinal mushroom historically used in Asian countries to treat a wide variety of diseases and prolong life. In the last years, G. lucidum has been internationally recognized as an effective adjuvant in cancer treatment. Among active components, the most recent research indicates that polysaccharides modulate the immune response favoring the recovery from toxicity of chemo and radiotherapy while triterpenes are cytotoxic to tumoral cells mainly by altering gene expression. Beyond this body of evidence on the efficacy of G. lucidum in cancer treatment, it is not yet understood whether these extracts exert the same mechanisms of action than current antitumoral drugs. Aim of the study In this study, we tested the DNA damaging potential of G. lucidum extracts by the β -galactosidase biochemical prophage induction assay (BIA) using doxorubicin, a DNA intercalating agent, as a positive control. This assay was traditionally used to screen microbial metabolites towards antitumoral agents. Here, we used this bacterial assay for the first time to assess DNA damage of herbal drugs. Results After a bioguided assay, only a purified fraction of G. lucidum containing a mixture of C16 and C18:1 fatty acids exerted weak activity which could not be attributed to direct interaction with DNA. At the same concentrations, the induction observed for doxorubicin was clearly contrasting. Conclusions The micro BIA assay could be successfully used to demonstrate differences in cellular effects between G. lucidum extracts and doxorubicin. These results showed that G. lucidum extracts display weak DNA damaging potential. Since DNA injury promotes aging and cancer, our results substantiate the traditional use of this mushroom to prolong life. [ABSTRACT FROM AUTHOR]

Abstract: We describe new plasmids that facilitate the rapid conversion of lacZ fusions to gfp transcriptional fusions in bacteria. The exchange is based on a double recombination between lacZ sequences in a suicide vector and the recipient chromosome. The suicide vector is a mobilizable, conditionally replicative plasmid that contains the gene for gfp with flanking lacZ homology and is derived from a broad host range plasmid that has been successfully used in a wide range of bacterial species. The technique was used to convert lacZ reporter fusions to gfp fusions in Escherichia coli, Bordetella bronchiseptica and Agrobacterium tumefaciens. Green fluorescent protein expression in the new recombinants reflected the beta galactosidase expression in the parent strains. GFP is particularly useful for rapid quantification of gene expression in real time and in single cells. As a demonstration of an application of this system, we studied the induction of virE transcription by the VirA/VirG two-component system in A. tumefaciens in response to various levels of phenolic inducer. Analysis of GFP fluorescence in single cells revealed that at intermediate levels of inducer the population of cells was remarkably heterogeneous. The tools described here will be useful for general studies of transcriptional regulation as well as for applications that require spatial and temporal identification of gene expression, such as in the study of biofilms, and interactions between bacteria and their environment. [Copyright &y& Elsevier]

The current investigation reports bactericidal and cytotoxicity evaluation of bio-enzymatically formulated nano‑gold (AuNPs) using physiologically significant enzyme β galactosidase. The AuNPs were characterized using spectroscopic and microscopic techniques. The anti-pathogenic efficacy of AuNPs as a potential drug was observed against five contagious bacterial strains of Escherichia coli , Staphylococcus aureus , group B Streptococcus , Acinetobacter baumannii and Klebsiella pneumonia. The estimation of minimum bactericidal concentration, minimum inhibitory concentration, scanning electron microscopy and fluorescence microscopy conclude enhanced bactericidal effects of green AuNPs. The cytotoxicity of AuNPs against human cervical (HeLa), breast (MCF-7) and liver (Hep 3B) cancer cells was evaluated. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was adopted to analyze antitumor potential of AuNPs. Cell nuclear morphology associated with apoptosis after treatment with NPs was assessed against MCF-7 cells through 4,6-diamidino-2-phenylindole staining. Apoptotic activity of AuNPs was determined against HeLa cells using annexin V/ propidium iodide double staining by flow cytometric analysis. The AuNPs exhibited excellent efficacy against these cell lines and future prospects of usage as potential nano-drugs and drug delivery vehicles. Unlabelled Image • Bio-enzymatic synthesis of nano‑gold by β galactosidase • Microscopic and biophysical characterization of AuNPs • Excellent inhibitory activity of AuNPs against five pathogenic bacterial cells • Enhanced cytotoxic effects of AuNPs against three different cancer cells [ABSTRACT FROM AUTHOR]