Low temperature desolventization: effect on physico-chemical, functional and structural properties of rice bran protein.

De-oiled rice bran is a good source of high-quality protein; however, the current practice of desolventization at high temperature (110-120 °C) denatures the protein, making its extraction difficult and uneconomical. The present study aims to investigate the effect of low temperature desolventization of de-oiled rice bran (LTDRB) on extraction, yield, and purity of protein and its comparison with protein obtained from high temperature desolventized de-oiled rice bran (HTDRB). The optimal conditions for preparation of protein from LTDRB were: extraction pH 11.00, extraction duration 52 min, and extraction temperature 58 °C resulting in an extraction efficiency, yield, and purity of 54.0, 7.23, and 78.70%, respectively. The LTDRB showed a positive impact on the color, solubility, foaming capacity and stability of protein whereas the absorption and emulsification properties were better for HTDRB protein. Significant decrease in enthalpy (ΔH) for denaturation was observed for LTDRB protein as compared to HTDRB protein. Scanning electron microscopy analysis revealed that HTDRB protein was more compact than LTDRB protein. LTDRB protein had smaller particle size distribution than HTDRB. Study suggested that low temperature desolventization can result in higher protein extraction with better physico-chemical, structural, and functional properties of protein obtained from DRB.

Recent advances in eco-friendly technology for decontamination of pulp and paper mill industrial effluent: a review.

The economic development of a country directly depends upon industries. But this economic development should not be at the cost of our natural environment. A substantial amount of water is spent during paper production, creating water scarcity and generating wastewater. Therefore, the Pollution Control Board classifies this industry into red category. Water is used in different papermaking stages such as debarking, pulping or bleaching, washing, and finishing. The wastewater thus generated contains lignin and xenobiotic compounds such as resin acids, chlorinated lignin, phenols, furans, dioxins, chlorophenols, adsorbable organic halogens (AOX), extractable organic halogens (EOCs), polychlorinated biphenyls, plasticizers, and polychlorinated dibenzodioxins. Nowadays, several microorganisms are used in the detoxification of these hazardous effluents. Researchers have found that microbial degradation is the most promising treatment method to remove high biological oxygen demand (BOD) and chemical oxygen demand (COD) from wastewater. Microorganisms also remove AOX toxicity, chlorinated compounds, suspended solids, color, lignin, derivatives, etc. from the pulp and paper mill effluents. But in the current scenario, mill effluents are known to deteriorate the environment and therefore it is highly desirable to deploy advanced technologies for effluent treatment. This review summarizes the eco-friendly advanced treatment technologies for effluents generated from pulp and paper mills.

Xylanolytic Enzymes in Pulp and Paper Industry: New Technologies and Perspectives.

The pulp and paper industry discharges massive amount of wastewater containing hazardous organochlorine compounds released during different processing stages. Therefore, some cost-effective and nonpolluting practices such as enzymatic treatments are required for the potential mitigation of effluents released in the environment. Various xylanolytic enzymes such as xylanases, laccases, cellulases and hemicellulases are used to hydrolyse raw materials in the paper manufacturing industry. These enzymes are used either individually or in combination, which has the efficient potential to be considered for bio-deinking and bio-bleaching components. They are highly dynamic, renewable, and high in specificity for enhancing paper quality. The xylanase act on the xylan and cellulases act on the cellulose fibers, and thus increase the bleaching efficacy of paper. Similarly, hemicellulase enzyme like endo-xylanases, arabinofuranosidase and ß-D-xylosidases have been described as functional properties towards the biodegradation of biomass. In contrast, laccase enzymes act as multi-copper oxidoreductases, bleaching the paper by the oxidation and reduction process. Laccases possess low redox potential compared to other enzymes, which need some redox mediators to catalyze. The enzymatic process can be affected by various factors such as pH, temperature, metal ions, incubation periods, etc. These factors can either increase or decrease the efficiency of the enzymes. This review draws attention to the xylanolytic enzyme-based advanced technologies for pulp bleaching in the paper industry.

Natural and grafted cyclotides in cancer therapy: An insight.

Cyclotides is a rapidly growing class of plant-derived cyclic peptides exhibiting several bioactivities with potential applications in the agricultural and pharmaceutical sectors. Both natural and grafted cyclotides have shown promise in cancer therapy. Approximately 70 natural cyclotides belonging to three plant families (Fabaceae, Rubiaceae, and Violaceae) have shown cytotoxicity against several cancer cell lines. Cyclotides exhibit considerable stability against thermal and enzymatic proteolysis, owing to their unique structure with knotted topology and head to tail cyclization. Further, their small size, high stability, oral bioavailability, and tolerance to amino acid substitution in structural loops make them an ideal platform for designing peptide-based drugs for cancer. Thus, cyclotides provide ideal scaffolds for bioactive epitope grafting and facilitating drug delivery in cancer treatment. Many anticancer linear peptides have been grafted in cysteine knotted cyclic framework of cyclotide for enhancing their cell permeability across cellular membranes, thereby improving their delivery and pharmacokinetics. The present review comprehensively discusses the distribution, toxicity, and anticancer bioactivity of natural cyclotides. Further, it systematically elaborates on the role and action of epitopes' into grafted cyclotides in targeting cancer. The review also encompasses related patents landscape study and future challenges in peptide-based cancer therapy.

Potency of Pfeiffer's Crystallization to Analyze Oral Leukoplakia and Squamous Cell Carcinoma.

OBJECTIVE: Oral cancer usually has an early precancerous stage before its actual malignant transformation. Although there are various approaches to diagnose early stages of cancer, yet there is one less explored, cost effective and simple technique known as crystallization test. The aim of the study was to reaffirm the effectiveness of Pfeiffer's crystallization test in screening oral leukoplakia and squamous cell carcinoma. METHODS: Fifty oral leukoplakia, sixty five oral squamous cell carcinoma and sixty healthy individuals participated in crystallization test. Single blood drop was used to perform the test and obtained crystal patterns were analysed. Cross tabulation and Chi-Square test was performed to find the frequency and association between the groups. Kruskal-Wallis H test and Mann Whitney U test was applied comparing mean transverse form. RESULTS: Sensitivity of crystallization test was 80% and 93.84% in oral leukoplakia and squamous cell carcinoma. Chi-Square analysis revealed highly significant transverse form between the study groups (p < 0.000). CONCLUSION: Crystallization test proves to be simple, reliable and minimal invasive diagnostic approach under strictly maintained physical conditions.

Microbial enzymes for deprivation of amino acid metabolism in malignant cells: biological strategy for cancer treatment.

Amino acid deprivation therapy (AADT) is emerging as a promising strategy for the development of novel therapeutics against cancer. This biological therapy relies upon the differences in the metabolism of cancer and normal cells. The rapid growth of tumors results in decreased expression of certain enzymes leading to auxotrophy for some specific amino acids. These auxotrophic tumors are targeted by amino acid-depleting enzymes. The depletion of amino acid selectively inhibits tumor growth as the normal cells can synthesize amino acids by their usual machinery. The enzymes used in AADT are mostly obtained from microbes for their easy availability. Microbial L-asparaginase is already approved by FDA for the treatment of acute lymphoblastic leukemia. Arginine deiminase and methionase are under clinical trials and the therapeutic potential of lysine oxidase, glutaminase and phenylalanine ammonia lyase is also being explored. The present review provides an overview of microbial amino acid depriving enzymes. Various attributes of these enzymes like structure, mode of action, production, formulations, and targeted cancers are discussed. The challenges faced and the combat strategies to establish AADT in standard cancer armamentarium are also reviewed.Key Points • Amino acid deprivation therapy is a potential therapy for auxotrophic tumors. • Microbial enzymes are used due to their ease of manipulation and high productivity. • Enzyme properties are improved by PEGylation, encapsulation, and genetic engineering. • AADT can be employed as combinational therapy for better containment of cancer.

Simultaneous laccase production and transformation of bisphenol-A and triclosan using Trametes versicolor.

New age micro-pollutants, bisphenol-A (BPA) and triclosan (TCA), known for their carcinogenic effects in living organisms can effectively be removed from water using laccase from Trametes versicolor. Laccase was produced from T. versicolor JSRK13 in both submerged and solid-state fermentation (SmF and SSF) conditions. In SmF, T. versicolor JSRK13 gave the maximum production of laccase on the 10th day with an activity of 22 U mL- 1, whereas, in SSF 185 U g- 1 of the enzyme was produced on the 17th day. Maximum production of laccase was observed with Parthenium as substrate. Parthenium, with a particle size of 3-5 mm having 60% moisture was found to be a suitable substrate for laccase production and simultaneous transformation (LPST) of BPA in a synergistic manner. A one-step concentration using 85% ammonium sulphate followed by dialysis was sufficient to give 6.7-fold purification of laccase from the crude culture filtrate. Transformation of BPA was achieved in both SmF and SSF conditions along with the production of laccase, whereas TCA was degraded with free enzyme only. Above 90% of BPA (55-5 mg L- 1) was degraded using the LPST strategy with HBT acting as a mediator in the reaction. LPST strategy did not work for TCA as it completely inhibits the growth of T. versicolor JSRK13. TCA was degraded up to 75% (1.5-0.375 mg L- 1) by the free enzyme. Our study of simultaneous laccase production and transformation proved to be efficacious in case of BPA. The results indicate that industrial and sewage wastewater containing BPA can potentially be treated with T. versicolor JSRK13 laccase. The described strategy can further be used to develop a bioprocess which can work both on solid and liquid wastes containing BPA.

Arginine-lowering enzymes against cancer: a technocommercial analysis through patent landscape.

INTRODUCTION: Rise in incidence of various cancers and growing adoption of biological therapy to avoid side effects of conventional cancer therapies is driving the growth of the cancer biotherapy market globally. One such therapy available for the treatment of certain tumors employs arginine-lowering enzymes (ALEs). Several patents have been filed in this technology domain, and many Phase I/II clinical trials of the ALEs especially arginine deiminase (ADI) are underway. AREAS COVERED: Patents and clinical trials in the domain of ALEs for the treatment of cancer were studied with an objective to understand technology trends, targeted areas, key players, and inventors involved. EXPERT OPINION: Amongst the various ALEs, ADI is the most promising enzyme for cancer therapy. ADI-based cancer therapy holds potential in treating liver, skin, lung, gastrointestinal, and blood cancer. ADI-PEG20 has proved to be very effective when used as a component of combination therapy in a first-line treatment. Polaris Group holds the worldwide rights for ADI-PEG20 and is the leading player in developing ADI as a therapeutic agent. Many clinical studies, especially in a combinatorial approach, are underway whose success will pave the way for ADI-PEG to the multimillion cancer market.

Laccase grafted membranes for advanced water filtration systems: a green approach to water purification technology.

PURPOSE: Conventional wastewater treatment technologies are not good enough to completely remove all endocrine disrupting compounds (EDCs) from the water. Membrane separation systems have emerged as an attractive alternative to conventional clarification processes for waste and drinking water. Coupling of a membrane separation process with an enzymatic reaction has opened up new avenues to further enhance the quality of water. This review article deliberates the feasibility of implementing enzymatic membrane reactors has been deliberated. MATERIALS AND METHODS: A comprehensive study of conventional water treatment technologies was carried out and their shortcomings were pointed out. Research findings from the leading groups working on enzyme grafted membrane based water purification were summarized. This review also comprehends the patent documents pertinent to the technology of enzyme grafted membranes for water purification. RESULTS: Immobilization of an enzyme on a membrane improves the performance of membrane filtration, and processes for the treatment of polluted water. Research has started exploring the potential for laccase enzymes because it can catalyze the oxidation of a wide range of substrates, structurally comparable to EDCs, by a radical-catalyzed reaction mechanism, with corresponding reduction of oxygen to water in an electron transfer process. Further, in the presence of certain mediators, the substrate range of laccases can be further enhanced to non-aromatic substrates. CONCLUSIONS: Removal of EDCs by laccase cross-linked enzyme aggregates in fixed-bed reactors or fluidized-bed reactors and laccase immobilized ultrafiltration (LIUF) membranes are proving their worth in water purification technology. The major operational issues with the use of LIUF membranes are enzyme instability in real wastewater and membrane fouling. In view of the above-stated characteristics, laccases are considered as the most promising enzyme for a greener and less expensive water purification technology.

Integrons in Enterobacteriaceae: diversity, distribution and epidemiology.

Integrons are versatile gene acquisition systems that allow efficient capturing of exogenous genes and ensure their expression. Various classes of integrons possessing a wide variety of gene cassettes are ubiquitously distributed in enteric bacteria worldwide. The epidemiology of integrons associated multidrug resistance in Enterobacteriaceae is rapidly evolving. In the past two decades, the incidence of integrons in enteric bacteria has increased drastically with evolution of multiple gene cassettes, novel gene arrangements and complex chromosomal integrons such as Salmonella genomic islands. This review focuses on the distribution, versatility, spread and global trends of integrons among important members of the Enterobacteriaceae, including Escherichia coli, Klebsiella, Shigella and Salmonella, which are known to cause infections globally. Such a comprehensive understanding of integron-associated antibiotic resistance, their role in the spread of such resistance traits and their clinical relevance especially with regard to each genus individually is paramount to contain the global spread of antibiotic resistance.

Applications of Trametes versicolor crude culture filtrates in detoxification of biomass pretreatment hydrolyzates.

Laccases have wide range of substrate specificity and find applications from pulp industry to waste water remediation. Laccases have also been used in combined pretreatment of biomass hydrolyzates to remove enzymatic and fermentation inhibitors. In this study, laccase production by Trametes versicolor strains isolated from different regions of the United States was induced using copper salts. T. versicolor crude culture filtrates (CCF), without any purification step, were tested for removal of model inhibitor compounds as well as in poplar and rice straw pretreatment hydrolyzates. Phenolic inhibitors were removed by 76% and 94% from the dilute acid hydrolyzates of rice straw and poplar, respectively, when incubated with the CCF for 12h, at room temperature. Xylo-oligosaccharide concentrations present in rice straw hydrolyzates were reduced by 64% when incubated with T. versicolor CCF. T. versicolor CCF could be a low cost technology for decreasing enzymatic and fermentation inhibitors.

Detoxification of sugarcane bagasse hydrolysate improves ethanol production by Candida shehatae NCIM 3501.

Sugarcane bagasse hydrolysis with 2.5% (v/v) HCl yielded 30.29g/L total reducing sugars along with various fermentation inhibitors such as furans, phenolics and acetic acid. The acid hydrolysate when treated with anion exchange resin brought about maximum reduction in furans (63.4%) and total phenolics (75.8%). Treatment of hydrolysate with activated charcoal caused 38.7% and 57.5% reduction in furans and total phenolics, respectively. Laccase reduced total phenolics (77.5%) without affecting furans and acetic acid content in the hydrolysate. Fermentation of these hydrolysates with Candida shehatae NCIM 3501 showed maximum ethanol yield (0.48g/g) from ion exchange treated hydrolysate, followed by activated charcoal (0.42g/g), laccase (0.37g/g), overliming (0.30g/g) and neutralized hydrolysate (0.22g/g).

Biochemical characterization and molecular evidence of a laccase from the bird's nest fungus Cyathus bulleri.

Cyathus bulleri, a bird's nest fungus, known to decolorize polymeric dye Poly R-478, was found to produce 8 U ml(-1) of laccase in malt extract broth. Laccase activity appeared as a single band on non-denaturing gel. Laccase was purified to homogeneity by anion exchange chromatography and gel filtration. The enzyme was a monomer with an apparent molecular mass of 60 kD, pI of 3.7 and was stable in the pH range of 2-6 with an optimum pH of 5.2. The optimal reaction temperature was 45 degrees C and the enzyme lost its activity above 70 degrees C. Enzyme could oxidize a broad range of various phenolic substrates. K(m) values for ABTS, 2,6-dimethoxyphenol, guaiacol, and ferulic acid were found to be 48.6, 56, 22, and 14 mM while K(cat) values were 204, 180, 95.6, and 5.2, respectively. It was completely inhibited by KCN, NaN(3), beta-mercaptoethanol, HgCl(2), and SDS, while EDTA had no effect on enzyme activity. The N-terminal amino acid sequence of C. bulleri laccase showed close homology to N-terminal sequences of laccase from other white-rot fungi. A 150 bp gene sequence encoding copper-binding domains I and II was most similar to the sequence encoding a laccase from Pycnoporus cinnabarinus with 74.8% level of similarity.