A critical review on the biotechnological potential of Brewers' waste: Challenges and future alternatives.

In order to comply with the stringent discharge guidelines issued by governmental organizations to protect the ecosystem, the substantial amounts of effluent and sturdy wastes produced by the beer brewing process need to be discarded or handled in the most affordable and secure manner. Huge quantities of waste material released with each brew bestow a significant opportunity for the brewing sector to move towards sustainability. The concept of circular economy and the development of technological advancements in brewery waste processing have spurred interest to valorize brewery waste for implementation in various sectors of medical and food science, industrial science, and many more intriguing fields. Biotechnological methods for valorizing brewery wastes are showing a path towards green chemistry and are feasible and advantageous to environment. The study unfolds most recent prospectus for brewery waste usage and discusses major challenges with brewery waste treatment and valorization and offers suggestions for further work.

Inhibition of respiratory syncytial virus by Daclatasvir and its derivatives: synthesis of computational derivatives as a new drug development.

The most common cause of respiratory tract illness in newborns and young children is the respiratory syncytial virus (RSV). There is no approved vaccination or specific antiviral medication for RSV infections. Here, an attempt has been made to explore the potential of currently marketed drugs as well as their probable derivatives to improve the possibility of developing stronger medications against RSV. From the 100 synthetic drug compounds library, the best drug molecule was identified through drug-likeness properties, toxicity, molecular docking and molecular dynamics simulations. Molecular Mechanics Generalized Born Surface Area (MM-GBSA) was also a method that was applied in this study. Daclatasvir showed the highest binding energy and appeared as the best drug to inhibit matrix protein and a fusion protein of RSV. Based on Daclatasvir, 40 computational derivatives were made. D28, D34 and D40 showed far better results than the actual drug. Changes in lipophilicity character increase the binding energy of derivatives. Molecular dynamic simulations showed their non-deviated, non-fluctuated and stable complex formation with target proteins. The high number of amino acid contacts throughout the trajectory increases the stability and effectiveness of derivatives. The key to producing a novel medicine to eradicate RSV is provided by derivatives. Daclatasvir will be employed as a potential RSV inhibitor up until that point.Communicated by Ramaswamy H. Sarma.

Potentiality of bioactive compounds as inhibitor of M protein and F protein function of human respiratory syncytial virus.

The human respiratory syncytial virus (RSV) creates a pandemic every year in several countries in the world. Lack of target therapeutics and absence of vaccines have prompted scientists to create novel vaccines or small chemical treatments against RSV's numerous targets. The matrix (M) protein and fusion (F) glycoprotein of RSV are well characterized and attractive drug targets. Five bioactive compounds from Alnus japonica (Thunb.) Steud. were taken into consideration as lead compounds. Drug-likeness characters of them showed the drugs are non-toxic and non-mutagenic and mostly lipophobic. Molecular docking reveals that all bioactive compounds have better binding and better inhibitory effect than ribavirin which is currently used against RSV. Praecoxin A appeared as the best lead compound between them. It creates 7 different types of bonds with amino acids of M protein and 5 different types of bonds with amino acids of F protein. Van der Waals interactions highly influenced the binding energies. Molecular dynamic simulations represent the non-deviated and less fluctuating nature of praecoxin A. Principal Component Analysis showed praecoxin A complex with RSV matrix protein is more stable than ribavirin complex. This study will help to develop a new drug to inhibit RSV. All ligands were minimized through semi-empirical PM3 process with MOPAC. Toxicity was tested by ProTox-II server. Molecular docking studies were carried out using AutoDock 4.2. Molecular dynamics simulations for 100 ns were carried out through GROMACS 5.12 MD and GROMOS96 43a1 force field. The graphs were produced by GROMACS's XMGrace program.

In silico comparative structural and compositional analysis of glycoproteins of RSV to study the nature of stability and transmissibility of RSV A.

The current scenario of COVID-19 makes us to think about the devastating diseases that kill so many people every year. Analysis of viral proteins contributes many things that are utterly useful in the evolution of therapeutic drugs and vaccines. In this study, sequence and structure of fusion glycoproteins and major surface glycoproteins of respiratory syncytial virus (RSV) were analysed to reveal the stability and transmission rate. RSV A has the highest abundance of aromatic residues. The Kyte-Doolittle scale indicates the hydrophilic nature of RSV A protein which leads to the higher transmission rate of this virus. Intra-protein interactions such as carbonyl interactions, cation-pi, and salt bridges were shown to be greater in RSV A compared to RSV B, which might lead to improved stability. This study discovered the presence of a network aromatic-sulphur interaction in viral proteins. Analysis of ligand binding pocket of RSV proteins indicated that drugs are performing better on RSV B than RSV A. It was also shown that increasing the number of tunnels in RSV A proteins boosts catalytic activity. This study will be helpful in drug discovery and vaccine development.

Purified cellulase-mediated simultaneous sugar utilization by Bacillus albus isolated from Similipal, Odisha, India.

Among 24 isolated cellulolytic bacteria from Similipal Biosphere Reserve, the most efficient isolate was recognized as a strain of Bacillus albus. This strain of B. albus was evaluated for cellulase production and the cellulase activity was measured in submerged fermentation using substrate carboxymethyl cellulose (CMC). Different nutritional (carbon, nitrogen, and metal-ion sources) and physical variables (pH, temperature, substrate concentration, and incubation time) during the growth of B. albus were optimized to obtain maximum cellulase activity. The highest cellulase activity of 5.79 U/mL for B. albus was observed at pH 6.75, temperature 37.5°C, CMC concentration 8.5 g/L, and 42 h incubation time. Further, supplementation of glucose as a subsidiary carbon source, yeast extract, peptone as nitrogen sources, and MgSO4 and MnSO4 as metal-ion sources enhance the cellulase activity of B. albus. The purified enzyme was reported to have a molecular weight of ∼54 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A zymogram analysis evidenced the cellulase activity of the purified enzyme fractions obtained from diethylaminoethyl cellulose chromatography. The purified cellulase was reported to have an optimum pH and temperature of 7.0°C and 50°C, respectively with a capacity of retaining its 60% residual activity within pH 6.0-8.0 and temperature 30-40°C, respectively. The metal ions, K+ and Na+ were the activators, while Pb2+ and Hg2+ were the inhibitors for the purified cellulase. The purified cellulase showed Km and Vmax values of 0.38 M and 8.19 U/mL, respectively, in presence of the substrate CMC and also simultaneous consumption of both hexose and pentose sugars.

Threat of respiratory syncytial virus infection knocking the door: a proposed potential drug candidate through molecular dynamics simulations, a future alternative.

The discovery of antiviral approaches to prevent or cure respiratory syncytial virus (RSV) infections is critical, particularly because RSV is one of the most common causes of infant respiratory problems. There is currently no approved vaccination available to treat RSV infections. FDA has approved the drug ribavirin, but it is not sufficient to treat RSV. This work aimed to find and study in silico anti-RSV drugs that target matrix protein and nucleoprotein. In this study, we have identified five drug candidates that had better binding energies than ribavirin. Garenoxacin appeared as top lead compounds between them. AutoDock Vina was used to execute molecular docking of a library of chosen chemicals. The high-score compound was then confirmed using the Maestro 12.3 module's molecular dynamics simulation and the binding energies derived using Prime/Molecular Mechanics Generalized Born Surface Area (Prime/MM-GBSA). Comparative molecular dynamics simulations revealed that garenoxacin has better stability and high residue contacts with high binding affinity than ribavirin. This study showed garenoxacin could prevent RSV infection better than ribavirin. In pursuing a more effective RSV control drug, additional research into these chemicals in vitro and in vivo is essential.

Biosynthesis and characterization of nanoparticles, its advantages, various aspects and risk assessment to maintain the sustainable agriculture: Emerging technology in modern era science.

The current review aims to gain knowledge on the biosynthesis and characterization of nanoparticles (NPs), their multifactorial role, and emerging trends of NPs utilization in modern science, particularly in sustainable agriculture, for increased yield to solve the food problem in the coming era. However, it is well known that an environment-friendly resource is in excessive demand, and green chemistry is an advanced and rising resource in exploring eco-friendly processes. Plant extracts or other resources can be utilized to synthesize different types of NPS. Hence NPs can be synthesized by organic or inorganic molecules. Inorganic molecules are hydrophilic, biocompatible, and highly steady compared to organic types. NPs occur in numerous chemical conformations ranging from amphiphilic molecules to metal oxides, from artificial polymers to bulky biomolecules. NPs structures can be examined by different approaches, i.e., Raman spectroscopy, optical spectroscopy, X-ray fluorescence, and solid-state NMR. Nano-agrochemical is a unification of nanotechnology and agro-chemicals, which has brought about the manufacture of nano-fertilizers, nano-pesticides, nano-herbicides, nano-insecticides, and nano-fungicides. NPs can also be utilized as an antimicrobial solution, but the mode of action for antibacterial NPs is poorly understood. Presently known mechanisms comprise the induction of oxidative stress, the release of metal ions, and non-oxidative stress. Multiple modes of action towards microbes would be needed in a similar bacterial cell for antibacterial resistance to develop. Finally, we visualize multidisciplinary cooperative methods will be essential to fill the information gap in nano-agrochemicals and drive toward the usage of green NPs in agriculture and plant science study.

Potential inhibitors of SARS-CoV-2 (COVID 19) proteases PLpro and Mpro/ 3CLpro: molecular docking and simulation studies of three pertinent medicinal plant natural components.

The rapid spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) - coronavirus disease 2019 (COVID-19) has raised a severe global public health issue and creates a pandemic situation. The present work aims to study the molecular -docking and dynamic of three pertinent medicinal plants i.e. Eurycoma harmandiana, Sophora flavescens and Andrographis paniculata phyto-compounds against SARS-COV-2 papain-like protease (PLpro) and main protease (Mpro)/3-chymotrypsin-like protease (3CLpro). The interaction of protein targets and ligands was performed through AutoDock-Vina visualized using PyMOL and BIOVIA-Discovery Studio 2020. Molecular docking with canthin-6-one 9-O-beta-glucopyranoside showed highest binding affinity and less binding energy with both PLpro and Mpro/3CLpro proteases and was subjected to molecular dynamic (MD) simulations for a period of 100ns. Stability of the protein-ligand complexes was evaluated by different analyses. The binding free energy calculated using MM-PBSA and the results showed that the molecule must have stable interactions with the protein binding site. ADMET analysis of the compounds suggested that it is having drug-like properties like high gastrointestinal (GI) absorption, no blood-brain barrier permeability and high lipophilicity. The outcome revealed that canthin-6-one 9-O-beta-glucopyranoside can be used as a potential natural drug against COVID-19 protease.

Rhizobacteria mediated seed bio-priming triggers the resistance and plant growth for sustainable crop production.

Advanced technologies are commonly used in modern agriculture to break the yield barriers and increase crop productivity. Seeds treated with plant growth-promoting rhizobacteria (PGPR) are an effective bio-priming method to introduce beneficial microbial inocula into the rhizosphere or soil. Bio-priming is a type of seed treatment that employs biological entities, which involves the hydration of seeds and inoculation with beneficial microorganisms. Mainly, the seed bio-priming technique improves the seed quality, germination, viability, vigor index, growth promotion, production, and subsequent disease resistance by enhancing the uniform speed of germination and production of others growth regulators. In the majority of cases, bacterial inoculants mostly PGPR are used for seed bio-priming, it is an ecologically comprehensive strategy that uses selected PGPR to promote plant growth by producing regulatory substances, enhancing uptake of nutrients, protecting seedlings/plants from seed or soil-borne pathogens. Bio-priming methods using PGPR inoculants are becoming more common in modern agriculture as an alternative to chemical treatments. They are more environmentally sustainable and safer for future agriculture apart from improving plants and soil health.

Discovery of Novel Cyclic Salt Bridge in Thermophilic Bacterial Protease and Study of its Sequence and Structure.

The plausible explanation behind the stability of thermophilic protein is still yet to be defined more clearly. Here, an in silico study has been undertaken by investigating the sequence and structure of protease from thermophilic (tPro) bacteria and mesophilic (mPro) bacteria. Results showed that charged and uncharged polar residues have higher abundance in tPro. In extreme environment, the tPro is stabilized by high number of isolated and network salt bridges. A novel cyclic salt bridge is also found in a structure of tPro. High number of metal ion-binding site also helps in protein stabilization of thermophilic protease. Aromatic-aromatic interactions also play a crucial role in tPro stabilization. Formation of long network aromatic-aromatic interactions also first time reported here. Finally, the present study provides a major insight with a newly identified cyclic salt bridge in the stability of the enzyme, which may be helpful for protein engineering. It is also used in industrial applications for human welfare.

Contributions of protein microenvironment in tannase industrial applicability: An in-silico comparative study of pathogenic and non-pathogenic bacterial tannase.

Tannase is an inducible industrially important enzyme, produced by several microorganisms. A large number of bacteria have reported as tannase producers; however, some of them are pathogenic in nature. Therefore, it is quite uncertain whether the application of these tannase enzymes from such pathogenic bacteria is suitable for industries and human welfare. Till date, there is no clear evidence regarding which group of bacteria (non-pathogenic or pathogenic) is better suited for their application in the edge of industries with particular reference to the food industry. The present study is following the findings of the above queries. In this study, a large number of tannase protein sequences have been retrieved from the databases, including both non-pathogenic and pathogenic bacterial species. Physiochemical and evolutionary properties of those sequences have been evaluated. Results have shown that non-pathogenic bacterial tannase possesses a high number of acidic and basic amino acid residues as compared to their pathogenic counterparts. The acidic and basic amino acid residues of tannase provide unique microenvironment to it. In the other hand, the numbers of disorder forming residues are higher in tannase sequences of pathogenic bacteria. The study of tannase microenvironment leads in the formation of salt bridges, which finally favoring the stability and proper functioning of tannase. This is the first report of such observation on tannase enzyme using in silico approach. Study of the microenvironment concept will be helpful in protein engineering.

Development of co-culture yeast fermentation for efficient production of biobutanol from rice straw: A useful insight in valorization of agro industrial residues.

Escalating environmental concerns and petroleum demands leads into the present study. In this investigation delignification of rice straw was optimized by NaOH and H2SO4 pretreatment using L16 Taguchi orthogonal array. NaOH pretreatment revealed higher delignification as compared to H2SO4 and; further subjected to separate enzymatic hydrolysis and co-fermentation (SHCF) using RSM as the SHCF demonstrated a maximum glucose and xylose yield of 575 and 205 mg/g. Further, butanol concentration of 4.32 g/L was achieved from 20 g/L of sugar loadings by co-culture of Saccharomyces cerevisiae and Pichia sp. at 72 h of incubation time which was 79.25% higher as compared to monocultures of Pichia sp. Scale-up experiments with higher sugar loadings (90 g/L) demonstrated a butanol concentration of 13.3 g/L. The release of amino acids in co-culture and monoculture systems demonstrated that the addition of S. cerevisiae promoted the butanol synthesis pathway which led to higher butanol concentration.

Effect of feed supplementation with biosynthesized silver nanoparticles using leaf extract of Morus indica L. V1 on Bombyx mori L. (Lepidoptera: Bombycidae).

Herein, we report the synthesis of silver nanoparticles (AgNPs) by a green route using the aqueous leaf extract of Morus indica L. V1. The synthesized AgNPs exhibited maximum UV-Vis absorbance at 460 nm due to surface plasmon resonance. The average diameter (~54 nm) of AgNPs was measured from HR-TEM analysis. EDX spectra also supported the formation of AgNPs, and negative zeta potential value (-14 mV) suggested its stability. Moreover, a shift in the carbonyl stretching (from 1639 cm-1 to 1630 cm-1) was noted in the FT-IR spectra of leaf extract after AgNPs synthesis which confirm the role of natural products present in leaves for the conversion of silver ions to AgNPs. The four bright circular rings (111), (200), (220) and (311) observed in the selected area electron diffraction pattern are the characteristic reflections of face centered cubic crystalline silver. LC-MS/MS study revealed the presence of phytochemicals in the leaf extract which is responsible for the reduction of silver ions. MTT assay was performed to investigate the cytotoxicity of AgNPs against two human cell lines, namely HepG2 and WRL-68. The antibacterial study revealed that MIC value of the synthesized AgNPs was 80 µg/ml against Escherichia coli K12 and Staphylococcus aureus (MTCC 96). Finally, the synthesized AgNPs at 10 µg/ml dosages showed beneficial effects on the survivability, body weights of the Bombyx mori L. larvae, pupae, cocoons and shells weights via enhancing the feed efficacy.

Comparative genomics of Mycobacterium reveals evolutionary trends of M. avium complex.

Mycobacterium is gram positive, slow growing, disease causing Actinobacteria. Beside potential pathogenic species, Mycobacterium also contains opportunistic pathogens as well as free living non-pathogenic species. Disease related various analyses on Mycobacterium tuberculosis are very widespread. However, genomic study of overall Mycobacterium species for understanding the selection pressure on genes as well as evolution of the organism is still illusive. MLSA and 16s rDNA based analysis has been generated for 241 Mycobacterium strains and a detailed analysis of codon and amino acid usage bias of mycobacterial genes, their functional analysis have been done. Further the evolutionary features of M. avium complex also have been revealed. Mycobacterial genes are moderately GC rich showed higher expression level in PPs and significant negative correlation with biosynthetic cost of proteins. Translational selection pressure was observed in mycobacterial genes. MAC showed close relationship with NPs and higher evolutionary rate in MAC revealed their constant evolving nature.

Enhanced reducing sugar production by saccharification of lignocellulosic biomass, Pennisetum species through cellulase from a newly isolated Aspergillus fumigatus.

A cellulose degrading fungus Aspergillus fumigatus (CWSF-7) isolated from decomposed lignocellulosic waste containing soil was found to produce high titer of cellulases. The optimum activity of CMCase and FPase were 1.9 U/mL and 0.9 U/mL respectively while the highest protein concentration was found to be 1.2 mg/mL. Saccharification of two Pennisetum grass varieties [dennanath (DG) and hybrid napier grass (HNG)] were optimized using partially purified CMCase and FPase in equal concentration, i.e. a ratios of 1:1 and further with addition of commercial xylanase using response surface methodology (RSM). The production of total reducing sugar (TRS) using isolated cellulase were 396.6 and 355.8 (mg/g), whereas further addition of xylanase had higher TRS titers of 478.7 and 483.3 (mg/g) for DG and HNG respectively as evident from HPLC analysis. Further, characterization of the enzyme saccharified DG and HNG by SEM and ATR-FTIR revealed efficient hydrolysis of cellulose and partially hydrolysis of hemicellulose.

Role of probiotic Lactobacillus fermentum KKL1 in the preparation of a rice based fermented beverage.

A dominant lactic acid bacteria, Lactobacillus fermentum KKL1 was isolated from an Indian rice based fermented beverage and its fermentative behavior on rice was evaluated. The isolate grown well in rice and decreased the pH, with an increase of total titratable acidity on account of high yield in lactic acid and acetic acid. The production of α-amylase and glucoamylase by the strain reached plateau on 1st and 2nd day of fermentation respectively. The accumulation of malto-oligosaccharides of different degrees of polymerization was also found highest on 4th day. Besides, phytase activity along with accumulation of free minerals also unremittingly increased throughout the fermentation. The fermented materials showed free radical scavenging activity against DPPH radicals. In-vitro characteristics revealed the suitability of the isolate as probiotic organism. The above profiling revealed that probiotic L. fermentum KKL1 have the significant impact in preparation of rice beer and improves its functional characteristics.

Microbial, saccharifying and antioxidant properties of an Indian rice based fermented beverage.

Haria, a popular rice based ethnic fermented beverage, is consumed as a staple food and refreshing drink by the vast number of Indian tribal people. In this study, the composition of microbial consortia and the occurrence of some important nutraceuticals during haria preparation were investigated. The quantities of moulds and yeasts were highest at 2nd day, and then declined, but, on the contrary, the quantity of Lactic Acid Bacteria and Bifidobacterium sp. increased concurrently during the course of fermentation. Accumulation of starch hydrolytic enzymes along with different types of malto-oligosaccharides like maltotetrose (26.18µg/gm), maltotriose (28.16µg/gm), and maltose (26.94µg/gm) were also noted. Furthermore, GC-MS analysis indicated the occurrence of pyranose derivatives in the fermented products. The fermented materials showed higher free radicals scavenging activity (82.54%, 4th day) against DPPH radicals. These studies clearly demonstrated that the microbial interaction during fermentation of rice makes it more nutritious, and most likely more beneficial for health.

Production of chitin and bioactive materials from Black tiger shrimp (Penaeus monodon) shell waste by the treatment of bacterial protease cocktail.

The main objective of this study was to obtain chitin in pure form from a new crustacean waste material for industrial applications. Black tiger shrimp shell wastes are a rich source of protein and valuable bioactive carbohydrate polymers such as chitin. After removal of carotenoid, Black tiger shrimp shell wastes (BTSHWs) were treated with chemicals and protease enzyme to extract chitin. Box-Behnken response surface methodology was applied to optimize the deproteinization process to obtain chitin. At optimal pH (8.82), temperature (50.05 °C), agitation speed (100.98 rpm), enzyme substrate ratio of 1:8 (wv-1) and 72 h of incubation with Paenibacillus woosongensis TKB2 crude protease cocktail, 80 % deproteinization was found along with 77.28 % recovery of chitin. The valuable oligopeptides were determined by MALDI-TOF analysis and analysis of adequate amount of free amino acids in protein hydrolysate from BTSHW, indicating a high nutritional value used for food, feed or as a nitrogen source in growth medium for microorganisms. The chitin obtained was compared with the commercial chitin using scanning electron microscopy, Fourier transform infrared spectrometer, X-ray diffraction and 13C CP/MAS-NMR. Chitin obtained from crude protease treatment showed comparable physicochemical and structural properties to those of the commercial chitin. The carotenoid obtained after treatment can be used for medicinal purpose.

Potentialities of newly isolated Bacillus subtilis and Lactobacillus sp for curd preparation and a comparative study of its physico-chemical parameters with other marketed curds.

Two Bacillus sp. were isolated from the local fermented milk and identified on the basis 16S rRNA sequence profile as Bacillus subtilis AKL1 and by biochemical process as Lactobacillus acidophilus AKL2. These isolates were used as fresh inoculums for curd preparation individually and in combinations. Different physico-chemical and therapeutic properties of the newly prepared curd were examined and compared with marketed local (sweet and sour) and branded (Mother Dairy and Thackar) curds. The total hydrolyzed peptides, free amino acids, lactic acid were significantly higher, whereas, total solid, ash content, syneresis and free reducing sugar were lower in the curd prepared by a mixture of AKL1 and AKL2 (0.5:0.5, v/v). The antioxidant activity against ABTS+, DPPH8, OH* and Fe3+ were also higher in the newly formulated curd. Polyphenols (85.5 microg/g), flavonoids (12.5 microg/g) and free aromatic amino acids contents were also higher in AKL1+AKL2. All these components prevent excess protein oxidation that was revealed by SDS-PAGE. The curd also exhibited potent antimicrobial activity against some entero-pathogens like Clostridium perfringens, Escherichia coli, Shigella dysentery, Vibrio cholerae and Staphylococcus aureus. It can be concluded that the combination of these Lactobacillus sp. will be a fruitful inoculum for the preparation of curd having better health promoting effects.