Improving the Growth and Productivity of Macrotyloma uniflorum Medicinal Plant by the Co-inoculation of P, Zn and K-Solubilizing Fungi Under Field Conditions.

The presence of small amount of soluble forms of Phosphorus (P), Potassium (K) and Zinc (Zn) in most soils is one of the limiting factors for agronomic crop production. The current study focuses on Macrotyloma uniflorum (horse gram or gahat), the most commonly cultivated crop in Uttarakhand. The current initiative and study were started, because there is a little information available on the impact of co-inoculation of beneficial fungi on crops in agricultural fields. Aspergillus niger K7 and Penicillium chrysogenum K4 were isolated and selected for the study on the basis of in vitro P, K and Zn-solubilizing activity. The solubilizing efficiency of K4 strain was 140% and K7 was 173.9% for P. However, the solubilizing efficiencies of K4 and K7 were 160% and 138.46% for Zn and 160% and 466% for K, respectively. The field trials were performed for two consecutive years, and growth and yield related parameters were measured for evaluation of the effect of P, K and Zn-solubilizing fungal strains on the crop. All the treatments showed a significant (P < 0.05) increase in growth and yield of M. uniflorum plants over uninoculated control; however, the best treatment was found to be soil inoculated with P. chrysogenum K4 + A. niger K7 in which the yield was enhanced by 71% over control. Thus, the co-inoculation of K4 and K7 strains showed a great potential to improve the growth and yield of plants. Both the fungal strains simultaneously solubilized three important nutritional elements in soil, which is a rare trait. Moreover, the capacity of these fungal strains to enhance the plant root nodulation and microbial count in soil makes the co-inoculation practice quite beneficial for sustainable agriculture.

Current trends in the development of soy-based foods containing probiotics and paving the path for soy-synbiotics.

In the world of highly processed foods, special attention is drawn to the nutrient composition and safety of consumed food products. Foods fortified with probiotic bacteria confer beneficial effects on human health and are categorized as functional foods. The salubrious activities of probiotics include the synthesis of vital bioactives, prevention of inflammatory diseases, anticancerous, hypocholesterolemic, and antidiarrheal effects. Soy foods are exemplary delivery vehicles for probiotics and prebiotics and there are diverse strategies to enhance their functionality like employing mixed culture fermentation, engineering probiotics, and incorporating prebiotics in fermented soy foods. High potential is ascribed to the concurrent use of probiotics and prebiotics in one product, termed as "synbiotics," which implicates synergy, in which a prebiotic ingredient particularly favors the growth and activity of a probiotic micro-organism. The insights on emended bioactive profile, metabolic role, and potential health benefits of advanced soy-based probiotic and synbiotic hold a promise which can be profitably implemented to meet consumer needs. This article reviews the available knowledge about strategies to enhance the nutraceutical potential, mechanisms, and health-promoting effects of advanced soy-based probiotics. Traditional fermentation merged with diverse strategies to improve the efficiency and health benefits of probiotics considered vital, are also discussed.

Production of ethanol, lipid and lactic acid from mixed agrowastes hydrolysate.

To combat the shortage of single agro-residue and overcome the problem of seasonal availability, it is beneficial to use mixture of lignocellulosic biomasses. In the present study, efforts were made to use mixed lignocellulosic biomass for production of bioethanol, along with microbial lipids and lactic acid. Upon enzymatic hydrolysis of mixed biomass at varied proportions it was observed that mixture of paddy straw and jute in the ratio 3:1 resulted in best sugar yield (41.50 g/L) at 10% substrate loading. Ethanolic fermentation of mixed substrate hydrolysate by thermotolerant yeast, Saccharomyces cerevisiae JRC6 resulted in 8.39 g/L of ethanol. To maintain sustainability and economic impact, oleaginous yeast (Trichosporon mycotoxinivorans S2) and lactic acid bacteria (Lactobacillus plantarum LP-9) were used for lipid production (14.5 g/L) and lactic acid production (11.08 g/L), respectively. Therefore, this study explored the potential of mixed lignocellulosic biomass to be exploited for production of various value-added products.

Bacterial volatile organic compounds as biopesticides, growth promoters and plant-defense elicitors: Current understanding and future scope.

Bacteria emit a large number of volatile organic compounds (VOCs) into the environment. VOCs are species-specific and their emission depends on environmental conditions, such as growth medium, pH, temperature, incubation time and interaction with other microorganisms. These VOCs can enhance plant growth, suppress pathogens and act as signaling molecules during plant-microorganism interactions. Some bacterial VOCs have been reported to show strong antimicrobial, nematicidal, pesticidal, plant defense, induced tolerance and plant-growth-promoting activities under controlled conditions. Commonly produced antifungal VOCs include dimethyl trisulfide, dimethyl disulfide, benzothiazole, nonane, decanone and 1-butanol. Species of Bacillus, Pseudomonas, Arthrobacter, Enterobacter and Burkholderia produce plant growth promoting VOCs, such as acetoin and 2,3-butenediol. These VOCs affect expression of genes involved in defense and development in plant species (i.e., Arabidopsis, tobacco, tomato, potato, millet and maize). VOCs are also implicated in altering pathogenesis-related genes, inducing systemic resistance, modulating plant metabolic pathways and acquiring nutrients. However, detailed mechanisms of action of VOCs need to be further explored. This review summarizes the bioactive VOCs produced by diverse bacterial species as an alternative to agrochemicals, their mechanism of action and challenges for employment of bacterial VOCs for sustainable agricultural practices. Future studies on technological improvements for bacterial VOCs application under greenhouse and open field conditions are warranted.

De-Oiled Citrus Peels as Feedstock for the Production of Pectin Oligosaccharides and Its Effect on Lactobacillus fermentum, Probiotic Source.

Following the extraction of essential oil, citrus (Mousambi, Kinnow, and Orange) peel wastes were used to produce pectin. The yield of essential oil and pectin was maximum in orange. Pectin was characterized by Fourier-transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD) spectroscopy. The degree of esterification (DE) and methoxyl content (MC) was maximum in orange whereas, the equivalent weight was maximum in Mousambi. A significant increase (61.8%) in the Lactobacillus fermentum population was observed with pectin as compared with sugar. Three sources followed the Orange > Kinnow > Mousambi trend as a prebiotic source. It was attributed to higher DE as well as higher MC. Enhancement in the bacterial population was in the range of 79.16-87.50%. The present work confirms the potential of pectin as a probiotic source for the enhancement of the bacterial population. Thus, it has a large scope for use in the food industry targeting a circular economy.

Bio-polysaccharide composites mediated degradation of polyaromatic hydrocarbons in a sandy soil using free and immobilized consortium of Kocuria rosea and Aspergillus sydowii.

Based on our previous study in minimal medium, Kocuria rosea and Aspergillus sydowii were identified as the best microbes for degradation of mixture of polyaromatic hydrocarbons (PAHs). The present study reports PAH degradation potential of these microbes in free and immobilized form. PAHs were extracted using QuEChERS-mediated process followed by quantification by high performance liquid chromatography. The microbial consortium of Kocuria rosea + Aspergillus sydowii was formulated in three bio-formulations, namely (i) bentonite-alginate composite beads; (ii) water dispersible granule composite using guar gum-nanobentonite; and (iii) composites of carboxymethyl cellulose-bentonite and were applied in PAH fortified (100 µg g-1) sandy loam soil. Results suggested that degradation data fitted well to first order kinetics as in most of the cases, the values of correlation coefficient (r) were > 0.95. The half-life (t1/2) values for PAHs in the uninoculated control soil were: naphthalene (10.43 d), fluorene (22.43 d), phenanthrene (24.64 d), anthracene (38.47 d), and pyrene (34.34 d). Inoculation of soil with free culture microbial consortium (without or with nutrient) and bio-formulation of degrading cultures enhanced degradation of all PAHs and half-life values were significantly reduced for each PAH: naphthalene (1.76-2.00 d), fluorene (2.52-6.65 d), phenanthrene (4.61-6.37 d), anthracene (9.01-12.22 d), and pyrene (10.98-15.55 d). Among different bio-formulations, guar gum-nanobentonite-based composite exhibited better efficacy for degradation of naphthalene, fluorene, phenanthrene, anthracene, and pyrene. The addition of microbial consortium in PAH fortified soil increased 16S rRNA gene copies of Alphaproteobacteria and Bacteroidetes, compared to the uninoculated, PAH-fortified control. The microbial functional gene assays showed that the gene copies of amoA, nirK, nirS, and anammox increased, suggesting nitrogen regulation in the PAH-fortified soil.

Potent γ-amino butyric acid producing psychobiotic Lactococcus lactis LP-68 from non-rhizospheric soil of Syzygium cumini (Black plum).

Gamma amino butyric acid (GABA) is a chemical messenger that plays a significant role in muscle relaxation and brain health. Certain lactic acid bacteria (LAB) produce significant levels of GABA and thus act as potential psychobiotic cultures. In the present study, LAB were isolated from non-rhizospheric soil sample of Syzygium cumini (Black plum). A total of 57 LAB were isolated on the basis of their morphological and acid producing characteristic on de Man Rogosa Sharpe (MRS) agar. Only seven isolates were found to produce GABA (0.09-1.13 gL-1) in MRS broth and were identified as Lactococcus. However, L. lactis LP-68 produced highest amount of GABA and was selected for further optimization of culture conditions (pH, temperature and MSG) by response surface methodology (RSM). The optimization resulted in approximately four-fold increase in GABA production (4.11 gL-1). The results indicate that the L. lactis LP-68 can be used as starter culture for production of GABA-enriched functional foods.

One-pot microbial bioconversion of wheat bran ferulic acid to biovanillin.

Due to growing consumer preference towards natural ingredients in food products, the production of flavors by microbial biotransformation of agrowastes provides an eco-friendly, cost-effective and sustainable pathway for biovanillin production. In the present study, biovanillin was produced by microbial biotransformation of ferulic acid (FA) using Streptomyces sp. ssr-198. The strain was able to grow in glucose medium supplemented with 1 g/L FA and produce 20.91 ± 1.11 mg/L vanillin within 96 h, along with 5.78 ± 0.13 mg/L vanillic acid in 144 h. Estimation of enzymes involved in FA degradation detected maximum feruloyl-CoA synthetase activity (1.21 ± 0.03 U/mg protein) at 96 h and maximum vanillin dehydrogenase activity (0.31 ± 0.008 U/mg protein) at 168 h, with small amounts of ferulic acid esterase activity (0.13 ± 0.002 U/mg protein) in the fermentation medium. Further, the glucose deficient production medium supplemented with 3 g/L of ferulic acid when inoculated with Streptomyces sp. ssr-198 (6% wet weight) produced maximum vanillin (685 ± 20.11 mg/L) within 72 h at 37 °C under agitation (150 rpm) and declined thereafter. Furthermore, in a one-pot experiment, wherein crude ferulic acid esterase (700 IU/g of substrate) from Enterococcus lactis SR1 was added into 10% w/v wheat bran (natural source of ferulic acid) based medium and was inoculated with 1% w/v of Streptomyces sp. ssr-198 resulted in maximum vanillin production (1.02 ± 0.02 mg/g of substrate) within 60 h of incubation. The study provides an insight into synergistic effect of using FAE of E. lactis SR1 and Streptomyces sp. ssr-198 for bioproduction of biovanillin using agro residues. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-03006-0.

Strain Improvement of Native Saccharomyces cerevisiae LN ITCC 8246 Strain Through Protoplast Fusion To Enhance Its Xylose Uptake.

Co-utilization of xylose and glucose and subsequent fermentation using Saccharomyces cerevisiae could enhance ethanol productivity. Directed engineering approaches have met with limited success due to interconnectivity of xylose metabolism with other intrinsic, hidden pathways. Therefore, random approaches like protoplast fusion were used to reprogram unidentified mechanisms. Saccharomyces cerevisiae LN, the best hexose fermenter, was fused with xylose fermenting Pichia stipitis NCIM 3498. Protoplasts prepared using glucanex were fused under electric impulse and fusants were selected using 10% ethanol and cycloheximide (50 ppm) markers. Two fusants, 1a.23 and 1a.30 showing fast growth on xylose and tolerance to 10% ethanol, were selected. Higher extracellular protein expression observed in fusants as compared to parents was corroborated by higher number of bands resolved by two-dimensional analysis. Overexpression of XYL1, XYL2, XKS, and XUT4 in fusants as compared to S. cerevisiae LN as observed by RT-PCR analysis was substantiated by higher specific activities of XR, XDH, and XKS enzymes in fusants. During lignocellulosic hydrolysate fermentation, fusants could utilize glucose faster than the parent P. stipitis NCIM 3498 and xylose consumption in fusants was higher than S. cerevisiae LN.

Restoration of heavy metal-contaminated soil and water through biosorbents: A review of current understanding and future challenges.

Heavy metal pollution in soil and water is a potential threat to human health as it renders food quality substandard. Different biosorbents such as microbial and agricultural biomass have been exploited for heavy metal immobilization in soil and sorptive removal in waters. Biosorption is an effective and sustainable method for heavy metal removal in soil and water, but the inherent challenges are to find cheap, selective, robust, and cost-effective bioadsorbents. Microbial and agricultural biomass and their modified forms such as nanocomposites and carbonaceous materials (viz., biochar, nanobiochar, biocarbon), might be useful for sequestration of heavy metals in soil via adsorption, ion exchange, complexation, precipitation, and enzymatic transformation mechanisms. In this review, potential biosorbents and their metal removal capacity in soil and water are discussed. The microbial adsorbents and modified composites of agricultural biomasses show improved performance, stability, reusability, and effectively immobilize heavy metals from soil and water. In the future, researchers may consider the modified composites, encapsulated biosorbents for soil and water remediation.

Cyanobacterial inoculation as resource conserving options for improving the soil nutrient availability and growth of maize genotypes.

Harnessing the benefits of plant-microbe interactions towards better nutrient mobilization and plant growth is an important challenge for agriculturists globally. In our investigation, the focus was towards analyzing the soil-plant-environment interactions of cyanobacteria-based formulations (Anabaena-Nostoc consortium, BF1-4 and Anabaena-Trichoderma biofilm, An-Tr) as inoculants for ten maize genotypes (V1-V10). Field experimentation using seeds treated with the formulations illustrated a significant increase of 1.3- to 3.8-fold in C-N mobilizing enzyme activities in plants, along with more than five- to six-fold higher values of nitrogen fixation in rhizosphere soil samples. An increase of 22-30% in soil available nitrogen was also observed at flag leaf stage, and 13-16% higher values were also recorded in terms of cob yield of V6 with An-Tr biofilm inoculation. Savings of 30 kg N ha-1 season-1 was indicative of the reduced environmental pollution, due to the use of microbial options. The use of cyanobacterial formulations also enhanced the economic, environmental and energy use efficiency. This was reflected as 37-41% reduced costs lowered GHG emission by 58-68 CO2 equivalents and input energy requirement by 3651-4296 MJ, over the uninoculated control, on hectare basis. This investigation highlights the superior performance of these formulations, not only in terms of efficient C-N mobilization in maize, but also making maize cultivation a more profitable enterprise. Such interactions can be explored as resource-conserving options, for future evaluation across ecologies and locations, particularly in the global climate change scenario.

Methane utilizing plant growth-promoting microbial diversity analysis of flooded paddy ecosystem of India.

Methane utilizing bacteria (MUB) are known to inhabit the flooded paddy ecosystem where they play an important role in regulating net methane (CH4) emission. We hypothesize that efficient MUB having plant growth-promoting (PGP) attributes can be used for developing novel bio-inoculant for flooded paddy ecosystem which might not only reduce methane emission but also assist in improving the plant growth parameters. Hence, soil and plant samples were collected from the phyllosphere, rhizosphere, and non-rhizosphere of five rice-growing regions of India at the tillering stage and investigated for efficient methane-oxidizing and PGP bacteria. Based on the monooxygenase activity and percent methane utilization on NMS medium with methane as the sole C source, 123 isolates were identified and grouped phylogenetically into 13 bacteria and 2 yeast genera. Among different regions, a significantly higher number of isolates were obtained from lowland flooded paddy ecosystems of Aduthurai (33.33%) followed by Ernakulum (20.33%) and Brahmaputra valley (19.51%) as compared to upland irrigated regions of Gaya (17.07%) and Varanasi (8.94%). Among sub-samples, a significantly higher number of isolates were found inhabiting the phyllosphere (58.54%) followed by non-rhizosphere (25.20%) and rhizosphere (15.45%). Significantly higher utilization of methane and PGP attributes were observed in 30 isolates belonging to genera Hyphomicrobium, Burkholderia, Methylobacterium, Paenibacillus, Pseudomonas, Rahnella, and Meyerozyma. M. oryzae MNL7 showed significantly better growth with 74.33% of CH4 utilization at the rate of 302.9 ± 5.58 and exhibited half-maximal growth rate, Ks of 1.92 ± 0.092 mg CH4 L-1. Besides the ability to utilize CH4, P. polymyxa MaAL70 possessed PGP attributes such as solubilization of P, K, and Zn, fixation of atmospheric N and production of indole acetic acid (IAA). Both these promising isolates can be explored in the future for developing novel biofertilizers for flooded paddies.

One-pot production of lactic acid from rice straw pretreated with ionic liquid.

Production of platform chemicals has been advocated as a sustainable option to tackle the problems associated with agro-waste management. In this report, for the first time, efforts were made to effectively produce second-generation lactic acid from rice straw pretreated with imidazolium ionic liquid [EMIM][OAc] and subsequently fermented with a promising Lactobacillus plantarum SKL-22 strain saccharified with a commercial cellulase enzyme. Medium optimization was carried out to enhance the lactic acid (LA) yield by response surface methodology. In a 5 L bioreactor, the process was further upscale, and a yield increment of 1.11% was observed. The process using rice straw as substrate led to a LA yield of 36.75 g/L from L. plantarum SKL-22 in a single pot bioprocess. Overall, the above finding has shown the ability of L. plantarum SKL-22 to produce LA from the hydrolysate of rice straw. This study presented a novel environmental-friendly method for LA production.

Overexpression and repression of key rate-limiting enzymes (acetyl CoA carboxylase and HMG reductase) to enhance fatty acid production from Rhodotorula mucilaginosa.

Implementing two-way strategies to enhance the lipid production in Rhodotorula mucilaginosa with the help of metabolic engineering was focused on the overexpression of acetyl coenzyme A carboxylase (ACC1 carboxylase) gene and repression of 3-hydroxy 3-methylglutaryl reductase (HMG-CoA reductase). Using an inducer (sodium citrate) and inhibitor (rosuvastatin), the amounts of biomass, lipid, and carotenoid were estimated. In the presence of inhibitor (200 mM), 62% higher lipid concentration was observed, while 44% enhancement was recorded when inducer (3 mM) was used. A combination of both inhibitor and inducer resulted in a 57% increase in lipid concentration by the oleaginous yeast. These results were again confirmed by real-time polymerase chain reaction by targeting the expression of the genes coding for ACC1 carboxylase and 13-fold increase was recorded in the presence of inducer as compared with control. This combined strategy (inducer and inhibitor use) has been reported for the first time as far as the best of our knowledge. The metabolic engineering strategies reported here will be a powerful approach for the enhanced commercial production of lipids.

Inulin from Pachyrhizus erosus root and its production intensification using evolutionary algorithm approach and response surface methodology.

Production of inulin from yam bean tubers by ultrasonic assisted extraction (UAE) was optimized by using response surface methodology (RSM) and genetic algorithms (GA). Yield of inulin was obtained between 11.97%-12.15% for UAE and 11.21%-11.38% for microwave assisted extraction (MAE) using both the methodologies, significantly higher than conventional method (9.9 %) using optimized conditions. Under such optimized condition, SEM image of root tissues before and extraction showed disruption and microfractures over surface. UAE provided a shade better purity of extracted inulin than other two techniques. Degree of polymerization in inulin was also recorded to be better, might be due lesser degradation during extraction. Significant prebiotic activity was recorded while evaluation using Lactobacillus fermentum and it was 36 % more than glucose treatment. Energy density by UAE was few fold lesser than MAE. Carbon emission was far more less in both these methods than the conventional one.

A simple downstream processing protocol for the recovery of lactic acid from the fermentation broth.

Lactic acid is one of the essential platform chemicals, and despite the availability of a range of downstream processes, its effective recovery is still elusive. A phase partitioning process using n-butanol and a chaotropic salt ammonium sulphate was developed to recover lactic acid from the fermentation broth of Lactobacillus pentosus SKL-18. During the optimization of various process parameters, the extraction medium's pH was found to be critical, with 2.5 being the best. The optimized process resulted in a lactic acid yield of 86% and found it to be 93% pure. The purity and characteristics of lactic acid were confirmed by FTIR and NMR spectra. This solvent-based extraction procedure is an economical and straightforward downstream process for purifying lactic acid produced from agro- and bakery-residues. The pure lactic acid can further be used for enzymatic synthesis of high value-added product PLA, a biodegradable and biocompatible plastics.

Efficient two-step lactic acid production from cassava biomass using thermostable enzyme cocktail and lactic acid bacteria: insights from hydrolysis optimization and proteomics analysis.

Lactic acid is an intermediate-volume specialty chemical, used in the production of biodegradable polymers and other chemicals. Although lactic acid production process is well established, however, the cost of production is very high. Therefore, in this study; starchy biomass (cassava) was hydrolyzed with in-house enzyme cocktail prepared from Aspergillus foetidus MTCC508 and Bacillus subtilis RA10. Process optimization using Taguchi experimental design helped to optimize the most effective ratio of fungal and bacterial amylase for effective saccharification of cassava. A higher sugar yield of 379.63 mg/gds was obtained under optimized conditions, using 30 U/gds of bacterial enzyme and 90 U/gds of the fungal enzyme at pH 4 within 48 h of saccharification. Among 11 lactic acid bacteria isolated, Lactobacillus fermentum S1A and Lactobacillus farraginis SS3A produced the highest amount of lactic acid 0.81 g/g and 0.77 g/g, respectively, from the cassava hydrolysate. The study proved the potential renewable source of cassava biomass as a source for fermentable sugars that can be fermented to lactic acid with high yield. In future, this cost-effective and environmental-friendly bioprocess can be upscaled for industrial lactic acid production.

A biorefinery approach for the production of ferulic acid from agroresidues through ferulic acid esterase of lactic acid bacteria.

Ferulic acid is a known precursor for vanillin production but the significance of agro waste as substrates for its extraction, in combination with microbes is a less explored option. Various lactic acid bacteria were screened for the production of ferulic acid esterase (FAE) and Enterococcus lactis SR1 was found to produce maximum FAE (7.54 ± 0.15 IU/ml) in the synthetic medium under submerged fermentation. To make the process cost effective, various lignocellulosic agroresidues were evaluated for the production of FAE from the bacterium. It was found that wheat bran serves as the best substrate for FAE production (4.18 ± 0.12 IU/ml) from E. lactis SR1. Further, optimization of fermentation conditions for FAE production from E. lactis SR1 using wheat bran as carbon source led to an increase in the enzyme production (7.09 ± 0.21 IU/ml) by 1.5 fold. The FAE produced was used alone or in combination with commercial holocellulase for biological release of FA from the tested agroresidues. The highest release of FA (mg/g) by enzymatic extraction occurred in sugarbeet pulp (2.56), followed by maize bran (1.45), wheat bran (1.39) and rice bran (0.87), when both the enzymes (FAE and holocellulase) were used together. Alkaline extraction and purification of ferulic acid (FA) from these agro residues also showed that sugarbeet pulp contains the highest amount of FA (5.5 mg/g) followed by maize bran (3.0 mg/g), wheat bran (2.8 mg/g) and rice bran (1.9 mg/g), similar to the trend obtained in biological/enzymatic extraction of FA from these residues. Furthermore, the substrates were found to release higher reducing sugars when both commercial holocellulase and FAE were used in combination than by the use of holocellulase alone. Thus, FAEs not only release FA but also enabled hemicellulase and cellulase to release more sugars from plant material.