Optimized process for the production of fungal peroxidases and efficient saccharification of pre-treated rice straw
Ambatkar N., Jadhav D.D., Nandi S., Kumbhar P., Kommoju P.-R.
Article, Bioresource Technology Reports, 2022, DOI Link
View abstract ⏷
Lignin is the most abundant natural source of aromatic polymers in plants. it prevents access to the underlying polymers of cellulose, hemicellulose/xylan for saccharification. Chemical pretreatment methods to remove this lignin barrier have their limitations such as corrosion of equipment, high capital costs, and environmental pollution due to emission of greenhouse gases. As an alternative, lignin modifying enzymes (LME) are being widely tested for delignification. While laccases are proven to break the lignin structure by attacking the phenolic bonds, additional enzymes to break bonds between non-phenolic units are needed. These reactions are commonly mediated by peroxidases such as Lignin peroxidases (LiP) and Versatile peroxidases (VP). Very limited data is available related to production of these very important peroxidases. In the current study, we present efficient production of these peroxidases from a white-rot fungi Pleurotus ostreatus and an optimized pre-treatment process showing an improvement of 20–25% cellulose to glucose conversion efficiency.
Functional screening and adaptation of fungal cultures to industrial media for improved delignification of rice straw
Ambatkar N., Jadhav D.D., Deshmukh A., Sattikar P., Wakade G., Nandi S., Kumbhar P., Kommoju P.-R.
Article, Biomass and Bioenergy, 2021, DOI Link
View abstract ⏷
Screening and development of industrial process for enzymatic delignification is an option for those industries who want to minimize acids and chlorine containing bleaches, thus minimizing the impact on environment. For commercial applications, it is important to screen fungi producing lignin modifying enzymes (LMEs) and adapting them to industrial media for submerged fermentations. Using functional plate assays and a full factorial design (FFD) approach, an optimal media composition for the highest expression of these LMEs has been achieved in this study. Two fungal strains (Trametes versicolor and Pleurotus ostreatus) were selected for evaluation. SDS PAGE and native gel analysis confirmed the results of screening and submerged culture media optimizations. When rice straw was added as inducer, secretion of additional enzymes other than laccases such as manganese peroxidase (MnP), lignin peroxidases (LiP), and versatile peroxidase (VP) in the cultures has been observed. This paper also demonstrates an improved process of enzymatic pretreatment resulting from a washing step introduced before saccharification (Scheme. 1). Hence, the current study proposes that removal of LMEs by washing helps in improvement of cellulase/hemicellulose depolymerization efficiency.
Regioselective and efficient enzymatic synthesis of antimicrobial andrographolide derivatives
Patil H.S., Jadhav D.D., Paul A., Mulani F.A., Karegaonkar S.J., Thulasiram H.V.
Article, Bioorganic and Medicinal Chemistry Letters, 2018, DOI Link
View abstract ⏷
Labdane diterpene andrographolide (1) is a major constituent of Andrographis paniculata and known to exhibit wide spectrum of biological activities. In this study, regioselective monoesters of (1) have been synthesized by using Amano lipase AK (Pseudomonas fluorescens) as a biocatalyst. Amano lipase AK was able to execute highly efficient esterification of hydroxyl group attached to C-14 carbon of (1) in presence of acyl donors. Among the various synthesized derivatives including two novel compounds such as andrographolide-14-propionate (3) and andrographolide-14-caproate (5) displayed antimicrobial activity against Staphylococcus aureus with low minimal inhibitory concentration (MIC) 4 µg/mL and 16 µg/mL respectively. Furthermore, they have shown low hemolysis activity at their respective MIC and increase in the permeability of the bacterial cell membrane as delineated by FITC uptake and SEM imaging studies.
Lipase catalyzed synthesis of antimicrobial andrographolide derivatives
Patil H.S., Jadhav D.D., Paul A., Mulani F.A., Karegaonkar S.J., Thulasiram H.V.
Data Paper, Data in Brief, 2018, DOI Link
View abstract ⏷
In this data article we describe screening of various lipases for the regioselective acylation of Andrographolide. Each lipase was screened with seven acyl donors. Amano lipase AK from Pseudomonas fluorescens was used for the synthesis of two new acylated andrographolide derivatives. Two new compounds, andrographolide-14-propionate and andrographolide-14-caproate were characterized by various spectral studies. These two derivatives showed more antimicrobial activity than andrographolide.
Oral feeding with polyunsaturated fatty acids fosters hematopoiesis and thrombopoiesis in healthy and bone marrow-transplanted mice
Limbkar K., Dhenge A., Jadhav D.D., Thulasiram H.V., Kale V., Limaye L.
Article, Journal of Nutritional Biochemistry, 2017, DOI Link
View abstract ⏷
Hematopoietic stem cells play the vital role of maintaining appropriate levels of cells in blood. Therefore, regulation of their fate is essential for their effective therapeutic use. Here we report the role of polyunsaturated fatty acids (PUFAs) in regulating hematopoiesis which has not been explored well so far. Mice were fed daily for 10 days with n-6/n-3 PUFAs, viz. linoleic acid (LA), arachidonic acid (AA), alpha-linolenic acid and docosahexanoic acid (DHA) in four separate test groups with phosphate-buffered saline fed mice as control set. The bone marrow cells of PUFA-fed mice showed a significantly higher hematopoiesis as assessed using side population, Lin-Sca-1+ckit+, colony-forming unit (CFU), long-term culture, CFU-spleen assay and engraftment potential as compared to the control set. Thrombopoiesis was also stimulated in PUFA-fed mice. A combination of DHA and AA was found to be more effective than when either was fed individually. Higher incorporation of PUFAs as well as products of their metabolism was observed in the bone marrow cells of PUFA-fed mice. A stimulation of the Wnt, CXCR4 and Notch1 pathways was observed in PUFA-fed mice. The clinical relevance of this study was evident when bone marrow-transplanted recipient mice, which were fed with PUFAs, showed higher engraftment of donor cells, suggesting that the bone marrow microenvironment may also be stimulated by feeding with PUFAs. These data indicate that oral administration of PUFAs in mice stimulates hematopoiesis and thrombopoiesis and could serve as a valuable supplemental therapy in situations of hematopoietic failure.
Data on the effect of oral feeding of Arachidonic acid or Docosahexanoic acid on haematopoiesis in mice
Limbkar K., Dhenge A., Jadhav D.D., Thulasiram H.V., Kale V., Limaye L.
Data Paper, Data in Brief, 2017, DOI Link
View abstract ⏷
Stem cells have peculiar property to self-renew and differentiate. It is important to control their fate in safe and effective ways for their therapeutic use. The mediators of essential polyunsaturated fatty acids (PUFAs) namely Arachidonic acid (AA) and Docosahexanoic acid (DHA) are known to play a role in haematopoiesis via various metabolic pathways [1]. However the direct effect of purified AA or DHA on haematopoiesis has not been well investigated yet. We have reported that oral administration of PUFAs enhanced haematopoiesis in mice [2]. Signaling Leukocyte Antigen Molecule (SLAM) (CD48−CD150+) phenotype consists of pure population of haematopoietic stem cells (HSCs). Herein we observed higher percentage of SLAM (CD48−CD150+) phenotype in the bone marrow (BM) cells of mice fed with AA or DHA compared to PBS fed control mice. Data from engraftment study depicts that BM from AA/DHA-fed mice showed higher absolute number of donor cells in recipient mice compared to control. The enhanced hematopoiesis observed in AA/DHA-fed mice was returned to normal when the mice were kept on normal diet for six weeks (after ten days of oral feeding). We confirmed GCMS (Gas Chromatography-Mass Spectroscopy) retention times of AA and DHA by co-injecting fatty acid extract from AA or DHA fed mice with purified AA or DHA standards respectively. Representative flow cytometry profile of Lin−Sca-1+c-kit+(LSK) cells showed higher expression of CXCR4 protein and ligands of Wnt, Notch1 signaling in BM of AA/DHA-fed mice.
Fungal mediated kinetic resolution of racemic acetates to (R)-alcohols using Fusarium proliferatum
Jadhav D.D., Patil H.S., Chaya P.S., Thulasiram H.V.
Article, Tetrahedron Letters, 2016, DOI Link
View abstract ⏷
Fungal mediated kinetic resolution of seven acyclic/aromatic acetates was achieved using Fusarium proliferatum to furnish (R)-alcohols in high enantiomeric excess (>95%). The kinetic resolution was established as one-pot two-step de-esterification/oxidation biocatalytic process. Further, the preparative scale synthesis of (R)-(+)-1-phenylethanol was accomplished through de-esterification/oxidation of (±)-1-phenylethyl acetate using the whole cell of F. proliferatum NCIM 1105.
Fungi mediated production and practical purification of (R)-(-)-3-quinuclidinol
Kolet S.P., Jadhav D.D., Priyadarshini B., Swarge B.N., Thulasiram H.V.
Article, Tetrahedron Letters, 2014, DOI Link
View abstract ⏷
A fungal system belonging to Mucoraceae family (Mucor piriformis) was explored for the asymmetric reduction of a prochiral ketone, 3-quinuclidinone (I) in an efficient manner to produce an important pharmaceutical precursor (R)-(-)-3-quinuclidinol (II) with ∼96% enantiomeric excess. The efficiency of the process was improved by developing a cation exchange resin (Amberlite IR-120) which assisted the purification of water soluble metabolite II from fermentation media.
Lipase mediated separation of triterpene structural isomers, α- And β-amyrin
Haldar S., Kale B.S., Jadhav D.D., Thulasiram H.V.
Article, Tetrahedron Letters, 2014, DOI Link
View abstract ⏷
Pentacyclic triterpenoids α- and β-amyrin possess a wide range of biological and pharmacological activities. High structural similarity between these two structural isomers makes their chromatographic separation an ineffective and tedious choice. In this study, Candida rugosa lipase catalyzed separation protocol for the isolation of individual isomers has been developed. In the presence of vinyl acetate as the acyl donor, Candida rugosa lipase carried out acetylation of β-amyrin more efficiently as compared to α-amyrin leading to a kinetic separation. The conditions of transesterification reaction were optimized systematically, which was utilized to separate α- and β-amyrin from a mixture obtained from the latex of Plumeria obtusa. © 2014 Elsevier Ltd. All rights reserved.
Differential induction, purification and characterization of cold active lipase from Yarrowia lipolytica NCIM 3639
Sathish Yadav K.N., Adsul M.G., Bastawde K.B., Jadhav D.D., Thulasiram H.V., Gokhale D.V.
Article, Bioresource Technology, 2011, DOI Link
View abstract ⏷
The production, purification and characterization of cold active lipases by Yarrowia lipolytica NCIM 3639 is described. The study presents a new finding of production of cell bound and extracellular lipase activities depending upon the substrate used for growth. The strain produced cell bound and extracellular lipase activity when grown on olive oil and Tween 80, respectively. The organism grew profusely at 20 °C and at initial pH of 5.5, producing maximum extracellular lipase. The purified lipase has a molecular mass of 400. kDa having 20 subunits forming a multimeric native protein. Further the enzyme displayed an optimum pH of 5.0 and optimum temperature of 25 °C. Peptide mass finger printing reveled that some peptides showed homologues sequence (42%) to Yarrowia lipolytica LIP8p. The studies on hydrolysis of racemic lavandulyl acetate revealed that extracellular and cell bound lipases show preference over the opposite antipodes of irregular monoterpene, lavandulyl acetate. © 2011 Elsevier Ltd.