Endophytic fungi of Panax sokpayensis produce bioactive ginsenoside Compound K in flask fermentation.

Endophytes of Panax have the potential to produce their host plant secondary metabolites, ginsenosides. Panax sokpayensis, an endemic traditional medicinal plant of the Sikkim Himalayas was explored for the isolation of endophytic fungi. In the present study, we have isolated 35 endophytic fungal cultures from the rhizome of P. sokpayensis and screened for ginsenosides production by HPLC by comparing the peak retention time with that of standard ginsenosides. The HPLC analysis revealed that out of 35 isolates, the mycelial extracts of four fungal endophytes (PSRF52, PSRF53, PSRF49 and PSRF58) exhibited peaks with a similar retention time of the standard ginsenoside, Compound K (CK). LC-ESI-MS/MS analysis led to the confirmation of ginsenoside CK production by the four fungal endophytes which showed a compound with m/z 639.6278, similar to that of standard ginsenoside CK with yield in potato dextrose broth flask fermentation ranging from 0.0019 to 0.0386 mg/g of mycelial mass in dry weight basis. The four prospective fungal endophyte isolates were identified as Thermothielavioides terrestris PSRF52, Aspergillus sp. PSRF49, Rutstroemiaceae sp. strain PSRF53, and Phaeosphaeriaceae sp. strain PSRF58 based on ITS sequencing. The present finding highlights the need for further study on growth optimization and other culture parameters to exploit the endophytes as an alternative source for ginsenoside CK production.

Potential of fermented foods and their metabolites in improving gut microbiota function and lowering gastrointestinal inflammation.

Foods prepared using microbial conversion of major and minor food components, which are otherwise known as fermented foods continue to impact human health. The live microorganisms and transformed metabolites can also have a deep influence on the gut microbiota, the multifaceted population of microorganisms dwelling inside the gut play a key role in wellbeing of an individual. The probiotic strains delivered through the consumption of fermented food and other bioactive components such as polyphenolic metabolites, bioactive peptides, short-chain fatty acids and others including those produced via gut microbiota mediated transformations have been proposed to balance the gut microbiota diversity and activity, and also to regulate the inflammation in the gut. However, little is known about such effects and only a handful of fermented foods have been explored to date. We herein review the recent knowledge on the dysbiotic gut microbiota linking to major gut inflammatory diseases. Also, evidences that fermented food consumption modulates the gut microbiota, and its impact on the gut inflammation and inflammatory diseases have been discussed. © 2024 Society of Chemical Industry.

Phytoconstituents from Urtica dioica (stinging nettle) of Sikkim Himalaya and their molecular docking interactions revealed their nutraceutical potential as α-amylase and α-glucosidase inhibitors.

In this study, antioxidative methanolic leaf extract (MeOH-SIS) of Urtica dioica was characterized for anti-diabetic activity. The extract was purified on a column to yield seven homogenous fractions (F1-F7) which were further determined for DPPH radical scavenging activity. MeOH-SIS and the fraction F1 (selected based on % yield and activity) were evaluated for their in vitro α-amylase and α-glucosidase inhibitory activity. The results showed inhibition of both enzymes in a dose dependent manner and F1 exhibited relatively higher inhibition than its mother extract MeOH-SIS. GC-MS analyses of both the extracts identified 24 major compounds among which 10 were previously described as bioactive compounds. Among all, 5 compounds demonstrated to have quality pharmacokinetics profiles and were examined for possible binding affinity against the active sites of α-amylase and α-glucosidase using molecular docking. The binding interaction of 2R-acetoxymethyl-1,3,3-trimethyl-4 T-(3-methyl-2-buten-1-yl)-1 T-cyclohexanol within the active sites of the target receptors was found to be significant among others, and can be developed as a potential inhibitor of α-amylase and α-glucosidase. The leaf extract can be utilized to develop food additive for the control and management of oxidative stress induced diabetes.

Assessment of the intrinsic bioremediation capacity of a complexly contaminated Yamuna River of India: a algae-specific approach.

Nearly 57 million people depend on Yamuna's water for their daily needs and agriculture. This is the first study of assessment of the Yamuna River for five major pollutants - Nitrate, Sulfate, Phosphate, Silicon, and Chloride, and the role of inhabitant algal species for phycoremediation. Water samples were collected from 11 different locations across three different seasons and it was found that the concentration of these pollutants varies in different locations and seasons. The concentration of Nitrate 392.93 mg/L at ITO Monsoon 2021, Phosphate 86.25 mg/L at Baghpat, Silicon 257.34 mg/L at Faridabad, Sulfate 2165.949 mg/L at ITO during winter 2020, and Chloride 4400.741 mg/L at Old bridge during Monsoon 2021 are found maximum. A significant variation (p < 0.05) in the concentrations of Nitrate, Sulfate, Phosphate, Silicon, and Chloride before and after treatment with microalgae was observed in water samples. All six algae significantly remove all the pollutants, and the maximum pollutants removed are Phosphate and Nitrite. Scenedesmus sp., removes the highest 99.21% Phosphate and 86.31% Nitrate, whereas 78.50% of Sulfate was removed by Klebsormidium sp. The highest 92.77% Silicon and 86.20% Chloride were removed by Oocystis sp. This finding suggests that out of six algae, Scenedesmus sp., in the Yamuna water has grown primarily at all the sites and reduces maximum pollutants. The outcomes from this study confirms that Yamuna River is highly contaminated at all the sites from these five major pollutants and algae are still survive in highly contaminated Yamuna water where no other plants are grown and phycoremediate the water bodies even in the presence of very high-stress condition. These algae can further be utilized for biotreatment of any contaminated water body.

This is the first study of assessment of five major pollutants and role of inhabitant algal species of Yamuna River for Phycoremediation.

Comparative analysis of the efficiency of seed protein profiles in assessing genetic variation and population structure among indigenous Manipur black rice cultivars.

BACKGROUND: The state of Manipur, North East India has distinct topology of hill and valley regions with vast agroclimatic variability, being considered as one of the centers of rice diversity. The indigenous Manipur black rice cultivars exhibit wide range of diversity in morphology, pericarp color, shape and size of grain, aroma, glutinous or non-glutinous features but remain less characterised. Many of these cultivars, such as those named Chakhao, are endowed with multiple health benefits due to high anthocyanins, and hold special importance for the local people. It is important to analyse the genetic diversity and population structure for this germplasm with unique allelic combinations to utilize in rice breeding programme. METHODS AND RESULTS: We characterized total soluble seed protein fractions to not only fingerprint the 45 indigenous black rice cultivars but assess their genetic relatedness. Cluster analyses generated mainly two groups, complemented by PCoA scatter plot ascertaining geographical distinction. The hill black rice were more diverse. The population structure analysis revealed seven subpopulations indicating high genetic variability. The 24 polymorphic bands were scored in the range of 127.8 to 10.3 kDa comprised of four protein fractions. Three polypeptide bands each were ascribed to known fractions of glutelins and prolamins, while one band each could be described for albumin and globulin fractions, besides other diagnostic bands. CONCLUSION: Some diverse cultivars were Amubi, Chedo Anal, Chipi Buh, Athebu, Poireton, BuPu Mui, Kotha Chahao II. These cultivars can be used in future black rice breeding programmes. This can further prevent genetic erosion and protect intellectual property rights.

Antioxidant Potential of Selected Wild Edible Leafy Vegetables of Sikkim Himalayan Region: Effects of Cooking Methods and Gastrointestinal Digestion on Activity.

Green leafy vegetables or GLVs are one of the main attractions in the local vegetable market and are widely consumed as the main course and side dish in the Sikkim Himalayan region (SHR). This study evaluated the total phenolic (TPC) and flavonoid contents (TFC) and antioxidant potential in different extracts such as methanolic (MeOH), ethyl acetate (EtOAC), and hexane extracts of selected GLVs followed by changes in the antioxidant activity on cooking and stimulated gastrointestinal (GI) digestion. The MeOH extracts of Urtica dioica L. (Sisnu), Nasturtium officinale W. T. Aiton (Simrayo), Diplazium esculentum Retz. Sw. (Ningro), and Chenopodium album L. (Bethu) were estimated to have higher TPC [22.73-45.84 µg gallic acid equivalent (GAE)/mg of extract]. In contrast, the plant extracts prepared using EtOAC (except for N. officinale, where TFC was found to be higher in hexane extract) were found to contain higher TFC (3.42-14.86 µg quercetin equivalent (QE)/mg of extract). The MeOH extracts also exhibited higher 2, 2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity (9.55-18.67 µg ascorbic acid equivalent (AAE)/mg of extract), total antioxidant activity (TAA) (0.27-0.32 mg AAE/mg of extract), and reducing power potential (RPP) (1.6-9.9 µg AAE/mg of extract). Among the test MeOH extracts, U. dioica demonstrated relatively higher antioxidant activities and was selected for cooking experiments followed by simulated GI digestion. The findings revealed that the loss of antioxidant activity was minimal in steam-cooked leaves (3.5% in 40 min) as compared to the boiled ones (18% in 10 min). The simulated GI (simulated salivary, gastric, and intestinal) digestion performed on raw, steam cooked, and boiled U. dioica leaves showed substantial enhancement of antioxidant properties (by 64.63%) through steam cooking in comparison to the raw leaves. Overall the study concludes that higher antioxidant properties can be achieved on the consumption of steam-cooked U. dioica leaves.

Production and characterization of bioactive peptides in novel functional soybean chhurpi produced using Lactobacillus delbrueckii WS4.

A novel soy chhurpi product was developed by fermentation of soymilk using proteolytic Lactobacillus delbrueckii strains isolated from traditional chhurpi production of Sikkim Himalaya. Soymilk fermentation by L. delbrueckii WS4 was associated with the hydrolysis of globulin proteins, with observed antioxidant, and ACE-inhibitory activity which further increased upon simulated in vitro gastrointestinal digestion. Peptidomics analysis of soy chhurpi and its gastrointestinal digest resulted in the identification of bioactive peptides with ACE-inhibitory and antioxidant properties. In silico antihypertensive property prediction followed by molecular docking study demonstrated strong binding affinity of selected peptides with ACE. The glycinin-derived peptide, SVIKPPTDE escaped gastrointestinal digestion and demonstrated strong non-bond interactions with ACE catalytic residues. QSAR models predicted an ACE-inhibitory IC50 of 21.29 µM for SVIKPPTDE. This is the first report on the production of novel functional soy chhurpi cheese using defined starter strains and the identification of bioactive peptides in undigested and gastrointestinal digested soy chhurpi.

Fermentation of black soybean with Bacillus spp. for the production of kinema: changes in antioxidant potential on fermentation and gastrointestinal digestion.

Black soybean was fermented with four different potential Bacillus spp., including Bacillus licheniformis K1G, Bacillus subtilis K2B, Bacillus amyloliquefaciens K2G and Bacillus subtilis K2M, isolated from kinema, a traditionally fermented soybean product of Sikkim. Enhancement of antioxidant activity was observed with DPPH radical scavenging activity, reducing power potential and total antioxidant activity in methanolic as well as water extracts. Overall antioxidant activities were found to be higher in fermented black soybean in comparison to yellow soybean, showing its potential for production of kinema. Further, black soybean fermented using different starter was subjected to gastrointestinal digestion using pepsin and pancreatin. Upon gastrointestinal digestion of fermented black soybean changes in antioxidant activity was observed that was found to be reliant on the species and strains applied for fermentation as starter culture. Among different starters used for fermentation, black soybean fermented using B. subtilis K2M had higher DPPH radical scavenging and reducing power activity on gastrointestinal digestion. This study concludes that B. subtilis K2M can be applied for fermentation of black soybean for production of kinema as well as bioactive protein hydrolysates. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13197-021-05144-y.

Draft genome sequence and potential identification of a biosurfactant from Brevibacterium casei strain LS14 an isolate from fresh water Loktak Lake.

This study reports the whole-genome sequencing and sequence analysis of a bacterial isolate Brevibacterium casei strain LS14, isolated from Loktak Lake, Imphal, India. The de novo assembled genome reported in this paper featured a size of 3,809,532 bp, has GC content of 68% and contains 3602 genomic features, including 3551 protein-coding genes, 46 tRNA and 5rRNA. A biosurfactant biosynthesis gene cluster in the genome of the isolated strain was identified using AntiSMASH online tool V3.0.5 and KAAS (KEGG Automatic Annotation Server). The presence of biosurfactant was demonstrated by drop collapse, oil displacement and emulsification index. Subsequent chemical characterization using FTIR and LC-MS analyses revealed surfactin and terpene containing biosurfactant moieties. Also, the presence of genes involved in terpenoid synthesis pathway in the genome sequence may account for biosurfactant terpenoid backbone, but genes for later-stage conversion of terpenoid to biosurfactant were not ascertained. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02867-9.

Nitrogen, phosphorus and high CO2 modulate photosynthesis, biomass and lipid production in the green alga Chlorella vulgaris.

Climate change could impact nutrient bioavailability in aquatic environment. To understand the interaction of nutrient bioavailability and elevated CO2, Chlorella vulgaris cells were grown in ambient air or 5% CO2 in different concentrations of nitrogen and phosphorus in a photobioreactor. The chlorophyll content, photosynthesis and respiration rates increased in 5% CO2 to support higher biomass production. The nutrient limitation in the growth media resulted in reduced photosynthetic rates of the algal cells and their PSI, PSII, and whole chain electron transport rates and biomass production. Conversely, their lipid content increased partly due to upregulation of expression of several lipid biosynthesis genes. The order of downregulation of photosynthesis and upregulation in lipid production due to nutrient limitation was in the order of N > P. The N-50 and 5% CO2 culture had only 10% reduction in biomass and 32% increase in lipids having 85% saturated fat required for efficient biofuel production. This growth condition is ideal for generation of biodiesel required to reduce the consumption of fossil fuel and combat global warming.

Production, characterization and molecular docking of antioxidant peptides from peptidome of kinema fermented with proteolytic Bacillus spp.

Kinema is an alkaline traditionally fermented soybean product popularly consumed in Sikkim Himalayan region. Kinema was prepared by soybean fermented with different species of Bacillus and analyzed for peptide content, antioxidant activity and consequence of gastrointestinal enzymes (pepsin and pancreatin) on the antioxidant effect. Antioxidant effect was enhanced during soybean fermentation using different starters, which further increased during gastrointestinal digestion. The peptides formed during soybean fermentation were analyzed using LC-MS/MS. Soybean fermented using different starters resulted in the production of some common peptides and a large number of unique peptides, which may affect the functional property of kinema. Peptides having antioxidative amino acids (histidine, phenylalanine, methionine, tryptophan and tyrosine) and significant GRAVY value were selected for their molecular interaction with myeloperoxidase (MPO), a key enzyme responsible for elevated oxidative stress. A peptide SEDDVFVIPAAYPF produced in kinema fermented using Bacillus licheniformis 1G had interaction with four out of five catalytic residues identified in MPO. Kinema prepared using specific starter can produce unique peptides responsible for specific health benefits.

Resource recovery through bioremediation of wastewaters and waste carbon by microalgae: a circular bioeconomy approach.

Microalgal biomass-based biofuels are a promising alternative to fossil fuels. Microalgal biofuels' major obstacles are the water and carbon sources for their cultivation and biomass harvest from the liquid medium. To date, an economically viable process is not available for algal based biofuels. The circular bioeconomy is an attractive concept for reuse, reduce, and recycle resources. The recovery of nutrients from waste and effluents by microalgae could significantly impact the escalating demands of energy and nutraceutical source to the growing population. Wastewaters from different sources are enriched with nutrients and carbon, and these resources can be recovered and utilized for the circular bioeconomy approach. However, the utilization of wastewaters and waste seems to be an essential strategy for mass cultivation of microalgae to minimizing freshwater consumption, carbon, nutrients cost, nitrogen, phosphorus removal, and other pollutants loads from wastewater and generating sustainable biomass for value addition for either biofuels or other chemicals. Hence, the amalgamation of wastewater treatment with the mass cultivation of microalgae improved the conventional treatment process and environmental impacts. This review provides complete information on the latest progress and developments of microalgae as potential biocatalyst for the remediation of wastewaters and waste carbon to recover resources through biomass with metabolites for various industrial applications and large-scale cultivation in wastewaters, and future perspectives are discussed.

Biotechnological approaches for the production of designer cheese with improved functionality.

Cheese is a product of ancient biotechnological practices, which has been revolutionized as a functional food product in many parts of the world. Bioactive compounds, such as peptides, polysaccharides, and fatty acids, have been identified in traditional cheese products, which demonstrate functional properties such as antihypertensive, antioxidant, immunomodulation, antidiabetic, and anticancer activities. Besides, cheese-making probiotic lactic acid bacteria (LAB) exert a positive impact on gut health, aiding in digestion, and improved nutrient absorption. Advancement in biotechnological research revealed the potential of metabolite production with prebiotics and bioactive functions in several strains of LAB, yeast, and filamentous fungi. The application of specific biocatalyst producing microbial strains enhances nutraceutical value, resulting in designer cheese products with multifarious health beneficial effects. This review summarizes the biotechnological approaches applied in designing cheese products with improved functional properties.

Metagenomics revealing molecular profiling of community structure and metabolic pathways in natural hot springs of the Sikkim Himalaya.

BACKGROUND: Himalaya is an ecologically pristine environment. The geo-tectonic activities have shaped various environmental niches with diverse microbial populations throughout the Himalayan biosphere region. Albeit, limited information is available in terms of molecular insights into the microbiome, including the uncultured microbes, of the Himalayan habitat. Hence, a vast majority of genomic resources are still under-explored from this region. Metagenome analysis has simplified the extensive in-depth exploration of diverse habitats. In the present study, the culture-independent whole metagenome sequencing methodology was employed for microbial diversity exploration and identification of genes involved in various metabolic pathways in two geothermal springs located at different altitudes in the Sikkim Himalaya. RESULTS: The two hot springs, Polok and Reshi, have distinct abiotic conditions. The average temperature of Polok and Reshi was recorded to be 62 °C and 43 °C, respectively. Both the aquatic habitats have alkaline geochemistry with pH in the range of 7-8. Community profile analysis revealed genomic evidence of plentiful bacteria, with a minute fraction of the archaeal population in hot water reservoirs of Polok and Reshi hot spring. Mesophilic microbes belonging to Proteobacteria and Firmicutes phyla were predominant at both the sites. Polok exhibited an extravagant representation of Chloroflexi, Deinococcus-Thermus, Aquificae, and Thermotogae. Metabolic potential analysis depicted orthologous genes associated with sulfur, nitrogen, and methane metabolism, contributed by the microflora in the hydrothermal system. The genomic information of many novel carbohydrate-transforming enzymes was deciphered in the metagenomic description. Further, the genomic capacity of antimicrobial biomolecules and antibiotic resistance were discerned. CONCLUSION: The study provided comprehensive molecular information about the microbial treasury as well as the metabolic features of the two geothermal sites. The thermal aquatic niches were found a potential bioresource of biocatalyst systems for biomass-processing. Overall, this study provides the whole metagenome based insights into the taxonomic and functional profiles of Polok and Reshi hot springs of the Sikkim Himalaya. The study generated a wealth of genomic data that can be explored for the discovery and characterization of novel genes encoding proteins of industrial importance.

Production and characterisation of lipase for application in detergent industry from a novel Pseudomonas helmanticensis HS6.

The aim of this study was to explore novel source of lipase from biodiversity hot spot region of Sikkim with activity at broad temperature range for application in detergent industry. Among the isolates, Pseudomonas helmanticensis HS6 showed activity at wide range of temperatures was selected for lipase production. Statistical optimisation for enhanced production of lipase resulted in enhancement of lipase activity from 2.3 to 179.3 U/mg. Lipase was purified resulting in 18.78 fold purification, 5.58% yield and high specific activity of 3368 U/mg. The partially purified lipase was found to be active in wide range of temperature (5-80 °C) and pH (6-9), showing optimum activity at 50 °C at pH 7. Peptide sequences on mass spectrometric analysis of purified lipase showed similarity to lipase family protein of three species of Pseudomonas. Both crude and purified lipase retained residual activity of 40-80% after 3 h of incubation with commercial detergents suggesting its application in detergent industry.

Molecular epidemiology of Japanese encephalitis virus in pig population of Odisha, Assam and Manipur states of India.

Japanese encephalitis virus (JEV) comes under the family Flaviviridae and genus flavivirus. Pigs act as reservoir and amplifying intermediate host for JEV. The current investigation was conducted to understand the prevalence of JEV infection in pigs in three different geographical sites in India (Odisha, Assam and Manipur). Total 857 serum samples were tested by ELISA and RT-PCR, while only RT-PCR was performed in case of 275 tonsils tissues for detection of JEV. It was observed that JEV prevalence was highest in Manipur (positive 39, 25.5% in serum and 10% in tonsil) but lower in Assam (positive 15, 3.8% in serum and 0% in tonsils) and Odisha (positive 7, 1.5% in serum and 3.7% in tonsils). Genotype III (GIII) of JEV was the dominant genotype. Further, analysis of E gene revealed sporadic mutations of S83G, H76P, E78Q, C55S, and S64W along with two consistent mutations V46S and V51I in GIII. Whereas, a single mutation S118N was observed in the GI strain. In conclusion, the high JE virus infection rate of pig in the current locations suggests the need for continuous surveillance of this virus in pigs which will ultimately help to adopt an effective control strategy to prevent the spread of JE infection to human.

Bioremediation and biomass production of microalgae cultivation in river watercontaminated with pharmaceutical effluent.

This work evaluated the potential of microalgae of Chlorella sp., SL7A, Chlorococcum sp., SL7B and Neochloris sp.,SK57 cultivated in river water contaminated with pharmaceutical effluent for biomass and lipid production. It has been observed that fast growing algae in this medium is Neochloris sp.SK57. Maximum biomass and lipid yield was obtained from Neochloris sp. SK57 (0.52 g/l) and Chlorococcum sp. SL7B (0.129 g/l)along with drycell weight of lipid was 28%.The increased in biomass and lipid in this media is could due to assimilation of organic nutrients and stress due to other components present in the river water. Fatty acid profile of algal biomass showed that saturated fatty acids production is enhanced in oils of Neochloris sp. SK57, and its suitability in food and fuel applications. Water quality of the river water was monitored before and after algal cultivation. Results showed that quality of river water was improved after algal cultivation.

De novo transcriptome assembly and analysis of Phragmites karka, an invasive halophyte, to study the mechanism of salinity stress tolerance.

With the rapidly deteriorating environmental conditions, the development of stress tolerant plants has become a priority for sustaining agricultural productivity. Therefore, studying the process of stress tolerance in naturally tolerant species hold significant promise. Phragmites karka is an invasive plant species found abundantly in tropical and sub tropical regions, fresh water regions and brackish marshy areas, such as river banks and lake shores. The plant possesses the ability to adapt and survive under conditions of high salinity. We subjected P. karka seedlings to salt stress and carried out whole transcriptome profiling of leaf and root tissues. Assessing the global transcriptome changes under salt stress resulted in the identification of several genes that are differentially regulated under stress conditions in root and leaf tissue. A total of 161,403 unigenes were assembled and used as a reference for digital gene expression analysis. A number of key metabolic pathways were found to be over-represented. Digital gene expression analysis was validated using qRT-PCR. In addition, a number of different transcription factor families including WRKY, MYB, CCCH, NAC etc. were differentially expressed under salinity stress. Our data will facilitate further characterisation of genes involved in salinity stress tolerance in P. karka. The DEGs from our results are potential candidates for understanding and engineering abiotic stress tolerance in plants.