Serratiopeptidase, a proteolytic enzyme serves as an important anti-inflammatory and analgesic medication. Present study reports the production and purification of extracellular serratiopeptidase ...from an endophyte, Serratia marcescens MES-4, isolated from Morus rubra. Purification of the enzyme by Ion exchange chromatography led to the specific activity of 13,030 U/mg protein of serratiopeptidase, showcasing about 3.1 fold enhanced activity. The catalytic domain of the purified serratiopeptidase, composed of Zn coordinated with three histidine residues (His 209, His 213, and His 219), along with glutamate (Glu 210) and tyrosine (Tyr 249). The molecular mass, as determined by SDS-PAGE was ∼51 kDa. The purified serratiopeptidase displayed optimal activity at pH 9.0, temperature 50°C. Kinetic studies revealed Vmax and Km values of 33,333 U/mL and 1.66 mg/mL, respectively. Further, optimized conditions for the production of serratiopeptidase by Taguchi design led to the productivity of 87 U/mL/h with 87.9 fold enhanced production as compared to the previous conditions.
•Purification and characterization of serratiopeptidase from the endophyte Serratia marcescens MES-4, isolated from Morus rubra.•Enhancement of serratiopeptidase activity by Taguchi L16 orthogonal array design.•Productivity of 87U/mL/h with 87.9 fold enhanced production was achieved as compared to the previous conditions.
Serratiopeptidase (EC 3.4.24.40), a proteolytic enzyme, is one of the most promising enzymes being used in biopharmaceutical industry. Mulberry phyllosphere, being an unexplored niche for exploration ...of protease production, was chosen for the present study. Protease producing bacteria were isolated from the tissues of mulberry plant as well as its rhizospheric soil. Two protease producing bacteria belonging to
Serratia
genus were found to be potential serratiopeptidase producers. Among them, the endophyte, i.e.,
Serratia marcescens
MES-4 presented 95 Units/mL activity, while the soil isolate i.e.,
Serratia marcescens
MRS-11 presented 156 Units/mL activity.
COVID-19 pandemic caused by the SARS-CoV-2 virus has led to a worldwide crisis. In view of emerging variants time to time, there is a pressing need of effective COVID-19 therapeutics. Setomimycin, a ...rare tetrahydroanthracene antibiotic, remained unexplored for its therapeutic uses. Herein, we report our investigations on the potential of setomimycin as COVID-19 therapeutic. Pure setomimycin was isolated from
Streptomyces
sp. strain RA-WS2 from NW Himalayan region followed by establishing in silico as well as in vitro anti-SARS-CoV-2 property of the compound against SARS-CoV-2 main protease (M
pro
). It was found that the compound targets M
pro
enzyme with an IC
50
value of 12.02 ± 0.046 μM. The molecular docking study revealed that the compound targets Glu166 residue of M
pro
enzyme, hence preventing dimerization of SARS-CoV-2 M
pro
monomer. Additionally, the compound also exhibited anti-inflammatory and anti-oxidant property, suggesting that setomimycin may be a viable option for application against COVID-19 infections.
Graphical abstract
•First report on the antagonistic potential of psychrotrophic fungus Trichoderma velutinum ACR-P1.•T. velutinum ACR-P1 is a producer of cell wall degrading enzymes (CWDEs).•Mycoparasitism supported ...by NRPs & CWDEs may contribute to its biocontrol potential.•Formulation of T. velutinum ACR-P1 may be developed for its use as biocontrol agent.
Trichoderma is a well explored genus for plethora of bioactivities due to their cosmopolitan existence, diverse metabolomics, reproductive and competitive efficacies. Mycoparasitism and antibiotics production seems to be the probable mechanisms underlying their use as biocontrol agents. The aim of this study is to investigate the antagonistic potential of a new psychrotrophic fungus Trichoderma velutinum ACR-P1 as a promising biocontrol agent against destructive phytopathogens with probable underlying mechanisms. Antagonistic potential of T. velutinum ACR-P1 against the important phytopathogens, that is, Fusarium oxysporum, Verticillium dahliae, Alternaria alternata and Colletotrichum capsici was demonstrated by the in vitro dual culturing experiments. Also, probable mechanism underlying antagonism was studied by investigating enzymatic cell wall hydrolytic potential of the strain and potential for production of secondary metabolites i.e. non-ribosomal peptides (NRPs). The putative strain ACR-P1 showed immense potential of inhibiting the growth of all the four test phytopathogens and its capability of producing cell wall degrading enzymes (CWDE) i.e. chitinase, cellulase and protease utilizing colloidal chitin, carboxymethyl cellulose and milk protein respectively as sole source of carbon and energy as illustrated by growth and agar well diffusion assays. Also, metabolic profiling firstly revealed the production of two groups of the NRPs by T. velutinum ACR-P1 as reported in our earlier study on the strain (Sharma et al., 2016). Mycoparasitism supported by NRPs and CWDEs substantiate their use as biocontrol agents. The psychrotrophic strain is endowed with immense potential to be included as a new candidate in the list of biocontrol agents (BCA). The potential antagonism against the important destructive phytopathogens signifies T. velutinum ACR-P1 as a BCA and can be exploited in fields to get rid of devastating impacts of these phytopathogens on the crops.
Verbenone is a natural monoterpene present as an essential component in rosemary oil from Rosmarinus officinalis, Verbena triphylla and Eucalyptus globule. Microbial biotransformation of ...monoterpenoids to value-added products has immense industrial potential. This study involves the biotransformation of (-)-verbenone to (-)-10-hydroxyverbenone by a fungus i.e. Talaromyces purpurogenus strain MRS-F13 in fermentation broth during the exponential phase. Biotransformation reaction has been successfully demonstrated at 15 L scale in the bioreactor with 70% conversion within 96 h. The anti-inflammatory activity investigations revealed that (-)-verbenone and its biotransformed product exhibited moderate inhibition of TNF-α and nitric oxide, whereas (-)-10-hydroxyverbenone presented an improved anti-oxidant activity.
Highlights
Talaromyces purpurogenus catalysed biotransformation of (-)-verbenone to (-)-10-hydroxyverbenone has been demonstrated.
Approximately 70% bioconversion was obtained in 96 h in submerged culture at 15L bioreactor scale.
Biotransformed product (-)-10-hydroxyverbenone has shown improved anti-inflammatory activity without cytotoxicity.
Background: COVID-19 pandemic has forced the human population to rethink over the lifestyle and food habits being followed by them. During the current scenario, when cases of more virulent new strain ...are emerging and specific treatment are still underway, we must look back to the Darvin’s theory of “Survival of the fittest” and in order to sustain the pandemic, we must follow the rule “Prevention is better than cure”. Traditional Medicinal System can bring possible solution including prevention and control of COVID-19. Humic substances have been used for antiviral activities, signifying that it has potential applications in the management and prevention of infectious diseases.
Summary: Present article is focused on the assessment of the naturally occurring humic substances, which are major constituents of Shilajit, being used in traditional system of medicine and can be used against for the prevention of COVID-19.
Key message: Humic substances not only improve the immune system, but also have anti-inflammatory, antioxidant and antiviral activities against human RNA viruses. Therefore, while the entire world is preparing to deal with alarming threat of community spread of COVID-19, Humic substances may play a vital role in improving the innate immunity for prevention and management of SARS-CoV-2 infection
Consistent production of bioactives from microbial sources remains a big challenge for fermentation based bio-processes. Setomimycin, a rare 9,9'-bianthrylanthracene antibiotic reported to be active ...against Gram positive bacteria i.e.
Staphyloccocus aureus
,
Bacillus subtilis
,
Bacillus cereus
, and
Mycobacterium smegmatis
, including mycobacteria is one of the least exploited antibiotic. Present work aims to enhance and maximize setomimycin production using One Factor at a Time (OFAT) approach, followed by Taguchi L9 orthogonal array (OA) design in 30L fermenter. Four most influential parameters, namely carbon source, nitrogen source, air and agitation were selected for optimization studies. The optimized production medium supplemented with 150 g/L glycerol and 7.5 g/L soyabean meal with an agitation rate of 100 RPM and air flow rate of 20 LPM (Liters Per Minute) resulted in 675 mg/L setomimycin production within 96–108 h of fermentation as compared to the initial production i.e. 40 mg/L. Thus, an overall enhancement of 16.8 folds was achieved in setomimycin production after optimization in 30L fermenter.
Graphical Abstract
Biosynthesis of bisaryl preanthraquinone antibiotics by various microorganisms differs in monomeric subunits as well as their dimerization positions leading to different configurations. The present ...study relates to the production of rare bisaryl anthraquinone antibiotics by a new Streptomyces strain isolated from Shivalik region of NW Himalayas. In vitro anticancer and anti-migratory effects of Setomimycin (9,9′ bisanthraquinone antibiotic) was seen with a significant reduction in the expression of both MEK as well as ERK pathways in a dose dependent manner at 6.5 μM & 8 μM concentration in HCT-116 and 5.5 μM & 7 μM concentration in MCF-7 cells. In vivo studies in aggressive orthotopic mouse mammary carcinoma model (4T1) demonstrated about 76% reduction of primary tumor weight and 90.5% reduction in the tumor volume within two weeks. In vivo pharmacokinetics study of setomimycin revealed that it can be rapidly absorbed with an adequate plasma exposure and half-life which can be linked to its in vivo efficacy.
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•Setomimycin, a rare bisaryl anthraquinones isolated from novel Streptomyces sp. strain from NW Himalayas.•An optima increase of par-4 was observed at 8 μM in HCT-116 cells and 7 μM in MCF-7 cells.•In-vivo studies in 4T1 mice model resulted in 90.5% reduction in the tumor volume within two weeks.•Setomimycin could absorbe rapidly (Tmax< 20 min) with a maximum plasma concentration level (Cmax>1 μM).
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