Biodegradation of DEP, DIBP, and BBP by a psychrotolerant Sphingobium yanoikuyae strain P4: Degradation potentiality and mechanism study.

Phthalic acid esters (PAEs) are human made chemicals widely used as plasticizers to enhance the flexibility of plastic products. Due to the lack of chemical bonding between phthalates and plastics, these materials can easily enter the environment. Deleterious effects caused by this chemo-pollutant have drawn the attention of the scientific community to remediate them from different ecosystem. In this context, many bacterial strains have been reported across different habitats and Sphingobium yanoikuyae strain P4 is among the few psychrotolerant bacterial species reported to biodegrade simple and complex phthalates. In the present study, biodegradation of three structurally different PAEs viz., diethyl phthalate (DEP), di-isobutyl phthalate (DIBP), and butyl benzyl phthalate (BBP) have been investigated by the strain P4. Quantitative analyses through High-performance liquid chromatography (HPLC) revealed that the bacterium completely degraded 1 g/L of DEP, DIBP, and BBP supplemented individually in minimal media pH 7.0 within 72, 54, and 120 h of incubation, respectively, at 28 °C and under shake culture condition (180 rpm). In addition, the strain could grow in minimal media supplemented individually with up to 3 g/L of DEP and 10.0 g/L of DIBP and BBP at 28 °C and pH 7.0. The strain also could grow in metabolites resulting from biodegradation of DEP, DIBP, and BBP, viz. n-butanol, isobutanol, butyric acid, ethanol, benzyl alcohol, benzoic acid, phthalic acid, and protocatechuic acid. Furthermore, phthalic acid and protocatechuic acid were also detected as degradation pathway metabolites of DEP and DIBP by HPLC, which gave an initial idea about the biodegradation pathway(s) of these phthalates.

Genomics assisted characterization of plant growth-promoting and metabolite producing psychrotolerant Himalayan Chryseobacterium cucumeris PCH239.

Here, we report the first complete genome of a psychrotolerant and yellow-pigmented rhizobacteria Chryseobacterium cucumeris PCH239. It was obtained from the rhizospheric soil of the Himalayan plant Bergenia ciliata. The genome consists of a single contig (5.098 Mb), 36.3% G + C content, and 4899 genes. The cold adaptation, stress response, and DNA repair genes promote survivability in a high-altitude environment. PCH239 grows in temperature (10-37 °C), pH (6.0-8.0), and NaCl (2.0%). The genome derived plant growth-promoting activities of siderophore production (siderophore units 53 ± 0.6), phosphate metabolism (PSI 5.0 ± 0.8), protease, indole acetic acid production (17.3 ± 0.5 µg/ml), and ammonia (2.89 ± 0.4 µmoles) were experimentally validated. Interestingly, PCH239 treatment of Arabidopsis seeds significantly enhances germination, primary, and hairy root growth. In contrast, Vigna radiata and Cicer arietinum seeds had healthy radicle and plumule elongation, suggesting varied plant growth-promotion effects. Our findings suggested the potential of PCH239 as a bio-fertilizer and biocontrol agent in the challenging conditions of cold and hilly regions.

Rahnella sikkimica sp. nov., a novel cold-tolerant bacterium isolated from the glacier of Sikkim Himalaya with plant growth-promoting properties.

The current study describes a novel species with the strain name ERMR1:05T isolated from the forefield soil of East Rathong Glacier in West Sikkim Himalaya (India). The isolate was facultatively anaerobic, gram-stain negative, non-spore-forming, rod-shaped, and oxidase negative. Whole-genome-based bacterial core gene phylogenetic analysis placed the strain in the genus Rahnella, well separated from Rouxiella spp. The digital DNA-DNA hybridisation and average nucleotide identity values between strain ERMR1:05T and other members of genus Rahnella were below the proposed thresholds for the species delineation. Based on these results, a new species, Rahnella sikkimica sp. nov., is proposed with strain ERMR1:05T (CIP 111636T, MTCC 12598T) as the type strain. The bacterium showed upregulation of cold-stress genes in cold conditions. Additionally, the genome analysis of the bacterium showed the presence of plant growth-promotion factors suggesting its role in crop improvement in cold hilly regions.

Microbial pigments: Learning from Himalayan perspective to industrial applications.

Pigments are an essential part of life on earth, ranging from microbes to plants and humans. The physiological and environmental cues induce microbes to produce a broad spectrum of pigments, giving them adaptation and survival advantages. Microbial pigments are of great interest due to their natural origin, diverse biological activities, and wide applications in the food, pharmaceutical, cosmetics, and textile industries. Despite noticeable research on pigment-producing microbes, commercial successes are scarce, primarily from higher, remote, and inaccessible Himalayan niches. Therefore, substantial bioprospection integrated with advanced biotechnological strategies is required to commercialize microbial pigments successfully. The current review elaborates on pigment-producing microbes from a Himalayan perspective, offering tremendous opportunities for industrial applications. Additionally, it illustrates the ecological significance of microbial pigments and emphasizes the current status and prospects of microbial pigments production above the test tube scale.

Green bioprocess for degradation of synthetic dyes mixture using consortium of laccase-producing bacteria from Himalayan niches.

Microbial remediation of synthetic dyes from industrial effluents offers a sustainable and eco-friendly alternative. Herein, laccase-producing bacteria were isolated from decaying wood niches in the Himalayan region. A bacterial consortium (BC-I) was developed to decolorize synthetic dyes cocktail of three major groups (azo, anthraquinone, and triphenylmethane). BC-I consisted of Klebsiella sp. PCH427, Enterobacter sp. PCH428, and Pseudomonas sp. PCH429 can decolorize 77% of 240 mg/L dyes cocktail in 44 h at 37 °C. BC-I works under wide pH (4.0-10.0), a high salt concentration (NaCl, 10%), and low nutrients. Further, FT-IR and LC-MS validated the dyes cocktail degradation and identified the degraded products. Additionally, phytotoxicity analysis of BC-I treated dyes cocktail significantly reduced the toxicity to Vigna radiata and Cicer arietinum compared to untreated dyes cocktail. The present study has simulated environmental challenges of acidic, alkaline, and saline industrial dyes effluents, which are significant to bioremediation.

Polyphenolic Profiling, Antioxidant, and Antimicrobial Activities Revealed the Quality and Adaptive Behavior of Viola Species, a Dietary Spice in the Himalayas.

BACKGROUND: Himalayan Viola species (Banksha) are traditionally important herbs with versatile therapeutic benefits such as antitussive, analgesic, antipyretic, antimalarial, anti-inflammatory, and anticancerous ones. The current investigation was focused on exploring polyphenolic profiles, antioxidant, and antimicrobial potentials of wild viola species at 15 gradient locations (375-1829 m). METHODS: Morphological, physiochemical, and proximate analyses were carried out as per WHO guidelines for plant drug standardization. Total polyphenolic and flavonoid content were carried out using gallic acid and rutin equivalent. UPLC-DAD was used to profile the targeted polyphenols (gallic acid, vanillic acid, syringic acid, p-coumaric acid, ferulic acid, rutin, quercetin, luteolin, caffeic acid, and epicatechin). Similarly, all samples were screened for antioxidant and antimicrobial activity. Statistical analysis was used to correlate polyphenolic and targeted activities to assess Viola species adaptation behavior patterns. RESULTS: Viola canescens (V. canescens) and Viola pilosa (V. pilosa) were found abundantly at their respective sites. Among flowers and leaves, flowers of V. canescens and V. pilosa showed higher total polyphenolic and flavonoid content (51.4 ± 1.13 mg GAE/g and 65.05 ± 0.85 mg RE/g, and 33.26 ± 0.62 mg GAE/g and 36.10 ± 1.41 mg RE/g, respectively). Furthermore, UPLC-DAD showed the uppermost content of p-coumaric acid in flowers and ferulic acid in leaves, while rutin was significant in both the tissues. CONCLUSIONS: The adaptive behavior of Viola species showed variability in morphological characters with the altitudes, while targeted polyphenols and activities were significant at mid-altitudes. This research helps in the selection of right chemotype for agrotechnological interventions and the development of nutraceutical products.

Metagenomic insights into Himalayan glacial and kettle lake sediments revealed microbial community structure, function, and stress adaptation strategies.

Glacial and kettle lakes in the high-altitude Himalayas are unique habitats with significant scope for microbial ecology. The present study provides insights into bacterial community structure and function of the sediments of two high-altitude lakes using 16S amplicon and whole-genome shotgun (WGS) metagenomics. Microbial communities in the sediments of Parvati kund (glacial lake) and Bhoot ground (kettle lake) majorly consist of bacteria and a small fraction of archaea and eukaryota. The bacterial population has an abundance of phyla Proteobacteria, Bacteroidetes, Acidobacteria, Actinobacteria, Firmicutes, and Verrucomicrobia. Despite the common phyla, the sediments from each lake have a distinct distribution of bacterial and archaeal taxa. The analysis of the WGS metagenomes at the functional level provides a broad picture of microbial community metabolism of key elements and suggested chemotrophs as the major primary producers. In addition, the findings also revealed that polyhydroxyalkanoates (PHA) are a crucial stress adaptation molecule. The abundance of PHA metabolism in Alpha- and Betaproteobacteria and less representation in other bacterial and archaeal classes in both metagenomes was disclosed. The metagenomic insights provided an incisive view of the microbiome from Himalayan lake's sediments. It has also opened the scope for further bioprospection from virgin Himalayan niches.

Genomic insights revealed physiological diversity and industrial potential for Glaciimonas sp. PCH181 isolated from Satrundi glacier in Pangi-Chamba Himalaya.

Himalayan niches provide unprecedented opportunities for finding novel microbes of commercial importance. The present study investigated the genome sequence of Glaciimonas sp. PCH181 isolated from the glacial stream of Indian trans-Himalaya. The draft genome sequence has six contigs with 5.3 Mb size, 51.1% G + C content, and possesses 4876 genes. Phylogenomic analysis revealed PCH181 as a putative novel bacterium in the genus Glaciimonas. Genomic insight showed Glaciimonas sp. PCH181 enriched with genes for diverse physiology, cold/stress adaptation, and industrial potential. The presence of genes for CO2 fixation and hydrogen metabolism suggested for chemolithoautotrophy. However, genes for sugars and organic acids usage showed heterotrophy and validated by physiological experiments. Genes for the metabolism of phenol (up to 500 ppm) and biosynthesis of polyhydroxyalkanoate (25% of dry cell mass) were also verified. Collectively, we present the first whole genome sequencing in the genus Glaciimonas, a taxonomically, physiologically, and industrially noteworthy bacterium.

A New Extracellular ß-Galactosidase Producing Kluyveromyces sp. PCH397 from Yak Milk and Its Applications for Lactose Hydrolysis and Prebiotics Synthesis.

ß-Galactosidase is a crucial glycoside hydrolase enzyme with potential applications in the dairy, food, and pharmaceutical industries. The enzyme is produced in the intracellular environment by bacteria and yeast. The present study reports yeast Kluyveromyces sp. PCH397 isolated from yak milk, which has displayed extracellular ß-galactosidase activity in cell-free supernatant through the growth phase. To investigate further, cell counting and methylene blue staining of culture collected at different growth stages were performed and suggested for possible autolysis or cell lysis, thereby releasing enzymes into the extracellular medium. The maximum enzyme production (9.94 ± 2.53U/ml) was achieved at 37 °C in a modified deMan, Rogosa, and Sharpe (MRS) medium supplemented with lactose (1.5%) as a carbon source. The enzyme showed activity at a wide temperature range (4-50 °C), maximum at 50 °C in neutral pH (7.0). In addition to the hydrolysis of lactose (5.0%), crude ß-galactosidase also synthesized vital prebiotics (i.e., lactulose and galacto-oligosaccharides (GOS)). Additionally, ß-fructofuranosidase (FFase) activity in the culture supernatant ensued the synthesis of a significant prebiotic, fructo-oligosaccharides (FOS). Hence, the unique features such as extracellular enzymes production, efficient lactose hydrolysis, and broad temperature functionality by yeast isolate PCH397 are of industrial relevance. In conclusion, the present study unrevealed for the first time, extracellular production of ß-galactosidase from a new yeast source and its applications in milk lactose hydrolysis and synthesis of valuable prebiotics of industrial importance. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12088-021-00955-1.

Posttranscriptional Regulation of tnaA by Protein-RNA Interaction Mediated by Ribosomal Protein L4 in Escherichia coli.

Escherichia coli ribosomal protein (r-protein) L4 has extraribosomal biological functions. Previously, we described L4 as inhibiting RNase E activity through protein-protein interactions. Here, we report that from stabilized transcripts regulated by L4-RNase E, mRNA levels of tnaA (encoding tryptophanase from the tnaCAB operon) increased upon ectopic L4 expression, whereas TnaA protein levels decreased. However, at nonpermissive temperatures (to inactivate RNase E), tnaA mRNA and protein levels both increased in an rne temperature-sensitive [rne(Ts)] mutant strain. Thus, L4 protein fine-tunes TnaA protein levels independently of its inhibition of RNase E. We demonstrate that ectopically expressed L4 binds with transcribed spacer RNA between tnaC and tnaA and downregulates TnaA translation. We found that deletion of the 5' or 3' half of the spacer compared to the wild type resulted in a similar reduction in TnaA translation in the presence of L4. In vitro binding of L4 to the tnaC-tnaA transcribed spacer RNA results in changes to its secondary structure. We reveal that during early stationary-phase bacterial growth, steady-state levels of tnaA mRNA increased but TnaA protein levels decreased. We further confirm that endogenous L4 binds to tnaC-tnaA transcribed spacer RNA in cells at early stationary phase. Our results reveal the novel function of L4 in fine-tuning TnaA protein levels during cell growth and demonstrate that r-protein L4 acts as a translation regulator outside the ribosome and its own operon.IMPORTANCE Some ribosomal proteins have extraribosomal functions in addition to ribosome translation function. The extraribosomal functions of several r-proteins control operon expression by binding to own-operon transcripts. Previously, we discovered a posttranscriptional, RNase E-dependent regulatory role for r-protein L4 in the stabilization of stress-responsive transcripts. Here, we found an additional extraribosomal function for L4 in regulating the tna operon by L4-intergenic spacer mRNA interactions. L4 binds to the transcribed spacer RNA between tnaC and tnaA and alters the structural conformation of the spacer RNA, thereby reducing the translation of TnaA. Our study establishes a previously unknown L4-mediated mechanism for regulating gene expression, suggesting that bacterial cells have multiple strategies for controlling levels of tryptophanase in response to varied cell growth conditions.

Biodegradation of organophosphorus pesticide chlorpyrifos by Sphingobacterium sp. C1B, a psychrotolerant bacterium isolated from apple orchard in Himachal Pradesh of India.

A psychrotolerant Sphingobacterium sp. was isolated from the apple orchard situated in the Kufri region of Shimla, Himachal Pradesh, India using an enrichment culture technique having chlorpyrifos (CP) as the sole source of carbon and energy. Based on biochemical characterization and 16S rRNA analysis, the strain was identified as Sphingobacterium sp. C1B. The bacterium C1B was able to degrade chlorpyrifos ≥ 42 ppm and ≥ 36 ppm within 14 days at 20 °C and 15 °C, respectively. The strain was also able to degrade chlorpyrifos ≤ 35 ppm at 28 °C within 14 days. The enzyme organophosphorus hydrolase might be responsible for the initial degradation of CP by the strain C1B. Based on the HPLC and GCMS analysis, a probable degradation pathway has been proposed, which followed the path from chlorpyrifos to 3,5,6-trichloro-2-pyridinol to benzene, 1,3-bis (1,1-dimethylethyl) and then entered into the TCA cycle. Our current study revealed that the bacterium C1B was found to be a useful strain for the degradation of pesticide chlorpyrifos in the cold climatic environment.

Identification of host factors limiting the overexpression of recombinant Cu, Zn superoxide dismutase in Escherichia coli.

OBJECTIVE: Superoxide dismutase (SOD) enzyme has implications in modulating the cell's redox state. The study aims to explore the host genetic factors that limit the heterologous expression of a thermostable SOD from Potentilla atrosanguinea (Pa-SOD) in E. coli. RESULTS: It was observed that the heterologous expression of Pa-SOD in E. coli did not exhibit any enhancement after 1 h of induction. This led to the alteration in cell morphology and an increase in the doubling time of E. coli cells expressing Pa-SOD. Label-free quantification and MALDI-TOF/TOF-MS/MS analysis suggested differential expression of 81 proteins, of which 77 proteins were found to be downregulated and 4 were found to be upregulated in Pa-SOD expressing cells as compared to uninduced E. coli cells. Functional analysis of downregulated proteins shows involvement in molecular function, biological process, and were the part of a cellular component. The STRING database revealed interaction of an essential autoregulatory protein, RNase E with other proteins involved in biosynthetic processes, protein biosynthesis and folding, and cell division. Further, validation of RNase E protein revealed upregulation of rne at transcript level and downregulation of RNase E at protein level as compared to uninduced cells. CONCLUSIONS: The observations suggested the operation of multifaceted mechanisms with a key role of RNase E that regulated the expression of Pa-SOD at the physiological and molecular level. Since Pa-SOD has commercial applications, identification and manipulation of these networked genetic factors could lead to improvement of host strain for large-scale production of biologically active Pa-SOD and other heterologous proteins.

Complete genome sequence of Pseudomonas frederiksbergensis ERDD5:01 revealed genetic bases for survivability at high altitude ecosystem and bioprospection potential.

Pseudomonas frederiksbergensis ERDD5:01 is a psychrotrophic bacteria isolated from the glacial stream flowing from East Rathong glacier in Sikkim Himalaya. The strain showed survivability at high altitude stress conditions like freezing, frequent freeze-thaw cycles, and UV-C radiations. The complete genome of 5,746,824 bp circular chromosome and a plasmid of 371,027 bp was sequenced to understand the genetic basis of its survival strategy. Multiple copies of cold-associated genes encoding cold active chaperons, general stress response, osmotic stress, oxidative stress, membrane/cell wall alteration, carbon storage/starvation and, DNA repair mechanisms supported its survivability at extreme cold and radiations corroborating with the bacterial physiological findings. The molecular cold adaptation analysis in comparison with the genome of 15 mesophilic Pseudomonas species revealed functional insight into the strategies of cold adaptation. The genomic data also revealed the presence of industrially important enzymes.

Diverse culturable bacterial communities with cellulolytic potential revealed from pristine habitat in Indian trans-Himalaya.

The Pangi-Chamba Himalaya (PCH) region is very pristine, unique, and virgin niche for bioresource exploration. In the current study, for the first time, the bacterial diversity of this region was investigated for potential cellulose degraders. A total of 454 pure bacterial isolates were obtained from diverse sites in the PCH region, and 111 isolates were further selected for 16S rDNA characterization based on ARDRA grouping. The identified bacteria belonged to 28 genera representing four phyla: Firmicutes, Proteobacteria, Actinobacteria, and Bacteroidetes. Pseudomonas was most abundant genus, followed by Bacillus, Geobacillus, Arthrobacter, Paenibacillus, and Flavobacterium. In addition, six putative novel bacteria (based on 16S rDNA sequence similarity) and thermophiles from non-thermogenic sites were also reported for the first time. Screening for cellulose degradation ability on carboxymethyl cellulose plates revealed that 70.92% of bacteria were cellulolytic. The current study reports diverse bacterial genera (Arthrobacter, Paenibacillus, Chryseobacterium, Pedobacter, Streptomyces, Agromyces, Flavobacterium, and Pseudomonas) with high capacity for cellulose hydrolysis and cellulolytic functionality at wide pH and temperature not previously reported in the literature. Diverse bacterial genera with high cellulolytic activity in broad pH and temperature range provide opportunity to develop a bioprocess for efficient pretreatment of lignocellulosic biomass, which is currently being investigated.

Antimicrobial Homoisoflavonoids from the Rhizomes of Polygonatum verticillatum.

Three homoisoflavonoids, including a new compound, 5,7-dihydroxy-3-(4-methoxybenzyl)-8-methyl chroman-4-one (1), together with two known compounds, 5,7-dihydroxy-3-(2-hydroxy-4-methoxybenzyl)-8-methylchroman-4-one (2) and 5,7-dihydroxy-3-(2-hydroxy-4-methoxybenzyl)-chroman-4-one (3), were isolated from the rhizomes of Polygonatum verticillatum (L.) All. (P. verticillatum). Isolated compounds were characterized on the basis of UV, FT-IR, ESI-MS, and 1D-, 2D-NMR data. Further, different extract fractions and pure compounds from Polygonatum verticillatum were screened for their antimicrobial potential. Among three pure compounds, compound 2 was found most potent with good zone of microbial growth inhibition as compared to the standards.

Cytomegalovirus infection in pregnant women and its association with bad obstetric outcomes in Northern India.

BACKGROUND: Cytomegalovirus (CMV) infection during pregnancy is far more complex than other infections, due to ability of the virus to be frequently reactivated during the child bearing age and may vertically transmitted to the developing fetus in spite of maternal immunity. Therefore, in the current study we determined the prevalence of CMV infection in pregnant women and tried to identify the role of maternal CMV infection in adverse pregnancy outcomes in Northern India. In this case-control study, 517 pregnant women, out of them 200 in case group and 317 in the control group. The overall 31.72% (164/517) cases were found with active CMV infection. CMV positivity (p=0.026) was significantly associated with bad obstetric history (75/200, 37.50%) compared to normal pregnancy (89/317, 28.07%). CMV infection was predominantly observed in age group 21-25 years. CMV positivity have been found to be significantly higher in women from rural area as compare to those from urban area (p=0.028). However, no significant difference has been observed in case of occupation, income, and haemoglobin level.

Biotechnological interventions for harnessing podophyllotoxin from plant and fungal species: current status, challenges, and opportunities for its commercialization.

Podophyllotoxin is an aryltetralin lignan synthesized in several plant species, which is used in chemotherapies for cancers and tumor treatment. More potent semisynthetic derivatives of podophyllotoxin such as etoposide and teniposide are being developed and evaluated for their efficacy. To meet the ever increasing pharmaceutical needs, species having podophyllotoxin are uprooted extensively leading to the endangered status of selective species mainly Sinopodophyllum hexandrum. This has necessitated bioprospection of podophyllotoxin from different plant species to escalate the strain on this endangered species. The conventional and non-conventional mode of propagation and bioprospection with the integration of biotechnological interventions could contribute to sustainable supply of podophyllotoxin from the available plant resources. This review article is focused on the understanding of different means of propagation, development of genomic information, and its implications for elucidating podophyllotoxin biosynthesis and metabolic engineering of pathways. In addition, various strategies for sustainable production of this valuable metabolite are also discussed, besides a critical evaluation of future challenges and opportunities for the commercialization of podophyllotoxin.

An outbreak of encephalitis associated with echovirus 19 in Uttar Pradesh, India, in 2011.

A sequence-independent single-primer amplification method and a modified enterovirus VP1 gene typing primer were used for identification of echovirus 19 and enterovirus 101, which remained undiagnosed by standard enterovirus molecular typing methods. Six different serotypes were identified during this study, with the predominance of ECV 19 (n = 20) followed by echovirus 21 (n = 3), EV 69 and EV 101 (n = 2 each), coxsackievirus B5 and ECV 27 (n = 1 each). To our knowledge, this is the first report of enteroviruses 69 and 101 in encephalitis cases in India.

Prevalence of arsenic exposure in population of Ballia district from drinking water and its correlation with blood arsenic level.

An investigation was carried out to ascertain the effect of arsenic in the blocks of Ballia district in Uttar Pradesh in the upper and middle Ganga plain, India. Analysis of 100 drinking water samples revealed that arsenic concentration was below 10 µg l⁻¹ in 60% samples, 10-50 µg l⁻¹ in 6%, 100 µg l⁻¹ in 24% and 200 µg l⁻¹ in 10% samples, respectively. The arsenic concentration in drinking water ranged from 12.8 to 132.2 µg l⁻¹. The depth of source of drinking water (10-60 m) was also found with a mean of 36.12 ± 13.61 µg l⁻¹ arsenic concentration. Observations revealed that at depth ranging from 10 to 20 m, the mean level of arsenic concentration was 17.398 ± 21.796 µg l⁻¹, while at 21 to 40 m depth As level was 39.685 ± 40.832 µg l⁻¹ and at 41 to 60 m As level was 46.89 ± 52.80 µg l⁻¹, respectively. These observations revealed a significant positive correlation (r = 0.716, t = 4.215, P < 0.05) between depth and arsenic concentration in drinking water. The age of water sources were ranged from zero to 30 years. The study indicates that the older sources of drinking water showed higher chance of contamination. Results showed that group 21-30 years having maximum arsenic concentration with mean value of 52.57 ± 53.79 µg l⁻¹. Correlation analysis also showed a significant positive correlation (r = 0.801, t = 5.66, P < 0.05) between age of drinking water sources and their respective arsenic concentration (µg l⁻¹). Arsenic concentration in blood with mean value 0.226 ± 0.177 µg dl⁻¹ significantly increased as compared to control. The blood arsenic content correlated significantly (r = 0.6823, t = 3.93, P < 0.05) with drinking water arsenic level and exposure time (r = 0.545, t = 3.101 & *P < 0.05) for populations residing in Ballia districts. Observations and correlation analysis revealed that individuals having depth of drinking water sources 20-30 m were less affected with arsenic exposure.

Biochemical characterization of glutaminase-free L-asparaginases from Himalayan Pseudomonas and Rahnella spp. for acrylamide mitigation.

L-asparaginase having low glutaminase activity is important in clinical and food applications. Herein, glutaminase-free L-asparaginase (type I) coding genes from Pseudomonas sp. PCH182 (Ps-ASNase I) and Rahnella sp. PCH162 (Rs-ASNase I) was amplified using gene-specific primers, cloned into a pET-47b(+) vector, and plasmids were transformed into Escherichia coli (E. coli). Further, affinity chromatography purified recombinant proteins to homogeneity with monomer sizes of ~37.0 kDa. Purified Ps-ASNase I and Rs-ASNase I were active at wide pHs and temperatures with optimum activity at 50 °C (492 ± 5 U/mg) and 37 °C (308 ± 4 U/mg), respectively. Kinetic constant Km and Vmax for L-asparagine (Asn) were 2.7 ± 0.06 mM and 526.31 ± 4.0 U/mg for Ps-ASNase I, and 4.43 ± 1.06 mM and 434.78 ± 4.0 U/mg for Rs-ASNase I. Circular dichroism study revealed 29.3 % and 24.12 % α-helix structures in Ps-ASNase I and Rs-ASNase I, respectively. Upon their evaluation to mitigate acrylamide formation, 43 % and 34 % acrylamide (AA) reduction were achieved after pre-treatment of raw potato slices, consistent with 65 % and 59 % Asn reduction for Ps-ASNase I and Rs-ASNase I, respectively. Current findings suggested the potential of less explored intracellular L-asparaginase in AA mitigation for food safety.