Browsing by Author "Jacob, Samuel"
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Item Open Access Arabinose as an overlooked sugar for microbial bioproduction of chemical building blocks(Taylor and Francis, 2023-11-06) Kumar, Vinod; Agrawal, Deepti; Bommareddy, Rajesh Reddy; Islam, M. Ahsanul; Jacob, Samuel; Balan, Venkatesh; Singh, Vijai; Thakur, Vijay Kumar; Navani, Naveen Kumar; Scrutton, Nigel S.The circular economy is anticipated to bring a disruptive transformation in manufacturing technologies. Robust and industrial scalable microbial strains that can simultaneously assimilate and valorize multiple carbon substrates are highly desirable, as waste bioresources contain substantial amounts of renewable and fermentable carbon, which is diverse. Lignocellulosic biomass (LCB) is identified as an inexhaustible and alternative resource to reduce global dependence on oil. Glucose, xylose, and arabinose are the major monomeric sugars in LCB. However, primary research has focused on the use of glucose. On the other hand, the valorization of pentose sugars, xylose, and arabinose, has been mainly overlooked, despite possible assimilation by vast microbial communities. The present review highlights the research efforts that have explicitly proven the suitability of arabinose as the starting feedstock for producing various chemical building blocks via biological routes. It begins by analyzing the availability of various arabinose-rich biorenewable sources that can serve as potential feedstocks for biorefineries. The subsequent section outlines the current understanding of arabinose metabolism, biochemical routes prevalent in prokaryotic and eukaryotic systems, and possible products that can be derived from this sugar. Further, currently, exemplar products from arabinose, including arabitol, 2,3-butanediol, 1,2,3-butanetriol, ethanol, lactic acid, and xylitol are discussed, which have been produced by native and non-native microbial strains using metabolic engineering and genome editing tools. The final section deals with the challenges and obstacles associated with arabinose-based production, followed by concluding remarks and prospects.Item Open Access Biological production and recovery of 2,3-butanediol using arabinose from sugar beet pulp by Enterobacter ludwigii(Elsevier, 2022-04-21) Narisetty, Vivek; Narisetty, Sudheera; Jacob, Samuel; Kumar, Deepak; Leeke, Gary A.; Chandel, Anuj Kumar; Singh, Vijai; Srivastava, Vimal Chandra; Kumar, VinodSugar beet pulp (SBP) is a major byproduct from the sugar industries and consists of >20% w/w arabinose. The current work evaluated the potential of Enterobacter ludwigii assimilating pure arabinose and arabinose rich hydrolysate from SBP pellets for 2,3-butanediol (BDO) production. The hydrolysate was obtained through dilute acid pretreatment (DAP) with sulphuric acid. The process was optimized for acid and solid loading to obtain a hydrolysate free from furan derivatives. The effect of different levels of substrate (10–60 g/L) using pure arabinose was conducted in shake flask experiments, followed by co-fermentation with small amounts of glucose and SBP hydrolysate. After flask cultivations, BDO fermentations were carried-out in a bench-top bioreactor in batch and fed-batch modes using pure arabinose as well as SBP hydrolysate. The fed-batch culture led to BDO production of 42.9 and 35.5 g/L from pure arabinose and SBP hydrolysate with conversion yields of 0.31 and 0.29 g/g, respectively. Finally, BDO accumulated on pure arabinose and SBP hydrolysate were recovered using an aqueous two-phase extraction system. The recovery yield of BDO accumulated on arabinose and hydrolysate was ∼97%. The work demonstrated the feasibility of using SBP as a suitable feedstock for manufacturing BDO.Item Open Access Bioprocessing of fermentable sugars derived from water hyacinth into microbial lipids and single cell proteins by oleaginous yeast Rhodosporidium toruloides NCIM 3547(Springer, 2021-10-09) Alankar, Senthilnathan Sri Laxma; Sajesh, Nithianandam; Rastogi, Shrestha; Sakhuja, Simar; Rajeswari, Gunasekaran; Kumar, Vinod; Chandel, Anuj Kumar; Jacob, SamuelIn this study, we employed microwave-acid pretreatment for water hyacinth (WH) to obtain liquid hydrolysate that contains sugars derived from holocellulosic components of biomass for further oleaginous yeast fermentation. In order to remove the inhibitors such as furans after acid treatment, detoxification of hydrolysate was done and we compared the efficiency of this step with non-detoxified hydrolysate towards capability of the Rhodosporidium toruloides NCIM 3547 (an oleaginous yeast) to produce microbial lipid and single cell protein. The results indicated that the reducing sugar concentration was found to be higher in non-detoxified hydrolysate (65.41 g/L) than detoxified one (59.18 g/L). When the non-detoxified liquid hydrolysate was supplemented with yeast extract as a complex organic source for R. toruloides, resulted in a maximum lipid yield of about 0.813 ± 0.041 (g/g) and 53.60 ± 2.68 (g/g) of single cell protein content with 0.038 g/L/d of protein productivity. Two kinetic models, hybrid Logistic-Monod and Luedeking-Piret, were employed to assess the microbial growth and the substrate utilization that were found to be in well agreement with the experimental data with a coefficient of determination (R2) value ranging from 0.95 to 0.99 thereby demonstrating the efficiency of the hydrolysate supplemented media. Furthermore, GC-MS analysis of transesterified lipids revealed the presence of various FAME (fatty acid methyl esters) and also the presence of increased levels of total saturated fatty acids (35.03%) advocates its high potential in biodiesel production. This study demonstrates the feasibility of sustainable valorization of WH-derived liquid hydrolysate towards a greener biorefinery framework.Item Open Access A concerted enzymatic de-structuring of lignocellulosic materials using a compost-derived microbial consortia favoring the consolidated pretreatment and bio-saccharification(Elsevier, 2024-01-13) Rajeswari, Gunasekaran; Kumar, Vinod; Jacob, SamuelThe robustness of microbial consortia isolated from compost habitat encompasses the complementary metabolism that aids in consolidated bioprocessing (CBP) of lignocellulosic biomass (LCB) by division of labor across the symbionts. Composting of organic waste is deemed to be an efficient way of carbon recycling, where the syntrophic microbial population exerts a concerted action of lignin and polysaccharide (hemicellulose and cellulose) component of plant biomass. The potential of this interrelated microorganism could be enhanced through adaptive laboratory evolution (ALE) with LCB for its desired functional capabilities. Therefore, in this study, microbial symbionts derived from organic compost was enriched on saw dust (SD) (woody biomass), aloe vera leaf rind (AVLR) (agro-industrial waste) and commercial filter paper (FP) (pure cellulose) through ALE under different conditions. Later, the efficacy of enriched consortium (EC) on consolidated pretreatment and bio-saccharification was determined based on substrate degradation, endo-enzymes profiling and fermentable sugar yield. Among the treatment sets, AVLR biomass treated with EC-5 has resulted in the higher degradation rate of lignin (47.01 ± 0.66%, w/w) and polysaccharides (45.87 ± 1.82%, w/w) with a total sugar yield of about 60.01 ± 4.24 mg/g. In addition, the extent of structural disintegration of substrate after EC-treatment was clearly deciphered by FTIR and XRD analysis. And the factors of Pearson correlation matrix reinforces the potency of EC-5 by exhibiting a strong positive correlation between AVLR degradation and the sugar release. Thus, a consortium based CBP could promote the feasibility of establishing a sustainable second generation biorefinery framework.Item Open Access Deep eutectic solvent pretreatment of water hyacinth for improved holocellulosic saccharification and fermentative co-production of xylitol and lipids using Rhodosporidium toruloides NCIM 3547(MDPI, 2022-10-31) Umai, Ramachandran Devasena; Jacob, Samuel; Kumar, VinodIn this study, delignification of water hyacinth (WH) using a mild ionic liquid-like chemical deep eutectic solvent (DES) synthesized using choline chloride and urea was conducted and the process parameters were optimized by Box–Behnken design (BBD)-based response surface methodology (RSM). From the results, a delignification of 64.32 ± 4.08% (w/w) was obtained under 1:12.5 (biomass:DES ratio), 4.63 h (time) and 87 °C (temperature). Further, a dilute sulphuric acid (2%, v/v) hydrolysis was carried out to destabilize the hemicellulose that resulted in 23.7 ± 0.50 g/L of xylose. Fermentation of the obtained xylose was carried out using a red oleaginous yeast, Rhodosporidium toruloides NCIM 3547, with free and Ca2+-alginate-immobilized cells for xylitol production under microaerophilic conditions and obtained yields of 4.73 ± 0.40 g/L (168 h) and 9.18 ± 0.10 g/L (packed bed reactor with a retention time of 18 h), respectively. Further, when the same fermentation was performed under aerobic conditions about 40.93 ± 0.73% lipid accumulation was observed with free cells. For saccharification, Aspergillus-niger-derived cellulase was used and this resulted in a yield of 27.45 ± 0.04 g/L of glucose. The glucose-enriched hydrolysate was supplemented for fermentation under nitrogen starved conditions from which 46.81 ± 2.60% (w/w) lipid content was obtained.Item Embargo An eclectic review on dicarboxylic acid production through yeast cell factories and its industrial prominence(Springer, 2024-04-20) Kayalvizhi, Ramalingam; Sanjana, Jayacumar; Jacob, Samuel; Kumar, VinodDicarboxylic acid (DCA) is a multifaceted chemical intermediate, recoursed to produce many industrially important products such as adhesives, plasticizers, lubricants, polymers, etc. To bypass the shortcomings of the chemical methods of synthesis of DCA and to reduce fossil fuel footprints, bio-based synthesis is gaining attention. In pursuit of an eco-friendly sustainable alternative method of DCA production, microbial cell factories, and renewable organic resources are gaining popularity. Among the plethora of microbial communities, yeast is being favored industrially compared to bacterial fermentation due to its hyperosmotic and low pH tolerance and flexibility for gene manipulations. By application of rapidly evolving genetic manipulation techniques, the bio-based DCA production could be made more precise and economical. To bridge the gap between supply and demand of DCA, many strategies are employed to improve the fermentation. This review briefly outlines the advancements in DCA production using yeast cell factories with the exemplification of strain improvement strategies.Item Open Access Enhanced 2,3-Butanediol production by mutant Enterobacter ludwigii using Brewers’ spent grain hydrolysate: process optimization for a pragmatic biorefinery loom(Elsevier, 2021-06-18) Amraoui, Yassin; Prabhu, Ashish A.; Narisetty, Vivek; Coulon, Frederic; Chandel, Anuj Kumar; Willoughby, Nicholas; Jacob, Samuel; Koutinas, Apostolis; Kumar, Vinod2,3-Butanediol (BDO) is a fossil-based versatile bulk chemical with a multitude of applications. BDO can also be synthesized using microbial cell factories harnessing renewable feedstocks. However, the high cost of the substrate via microbial route impedes commercial manufacturing of BDO. Therefore, identification of cheaper substrates could make bio-based BDO production more cost-competitive. Brewers’ spent grain (BSG), a major by-product of breweries, is an inexpensive source of fermentable sugars and proteins. In the present study, we have attempted the bioproduction of BDO by Enterobacter ludwigii using BSG as feedstock. A random E. ludwigii mutant obtained after treatment with ethyl methane sulfonate (EMS) resulted in a BDO titer (9.5 g/L), ~30% higher in comparison to the wild type strain with a yield of 0.48 gBDO/gGlucose approaching the theoretical yield of 0.50 gBDO/gGlucose. The enzymatic hydrolysis of microwave-assisted alkali pretreated BSG was optimized using the statistical Taguchi design. The BSG hydrolysis under optimal conditions (pH: 6.0; temperature: 50 °C; BSG: 10% w/v; enzyme loading: 2% v/v) resulted in a glucose yield of 0.25 gGlucose/gBiomass. The uncontrolled pH was found to be more beneficial for BDO accumulation from BSG hydrolysate in batch bioreactor cultivation as compared with controlled one. The fed-batch cultivation with forced pH fluctuations at an aeration rate of 2.0 vvm resulted in BDO accumulation of 118.5 g/L from glucose-rich BSG hydrolysate with the yield and productivity of 0.43 g/g and 1.65 g/L.h, respectively. To the best of our knowledge, this is the first study on BDO production from BSG.Item Open Access Fermentative valorisation of xylose-rich hemicellulosic hydrolysates from agricultural waste residues for lactic acid production under non-sterile conditions(Elsevier, 2023-05-18) Cox, Rylan; Narisetty, Vivek; Castro, Eulogio; Agrawal, Deepti; Jacob, Samuel; Kumar, Gopalakrishnan; Kumar, Deepak; Kumar, VinodLactic acid (LA) is a platform chemical with diverse industrial applications. Presently, commercial production of LA is dominated by microbial fermentation using sugary or starch-based feedstocks. Research pursuits emphasizing towards sustainable production of LA using non-edible and renewable feedstocks have accelerated the use of lignocellulosic biomass (LCB). The present study focuses on the valorisation of xylose derived from sugarcane bagasse (SCB) and olive pits (OP) through hydrothermal and dilute acid pretreatment, respectively. The xylose-rich hydrolysate obtained was used for LA production by homo-fermentative and thermophilic Bacillus coagulans DSM2314 strain under non-sterile conditions. The fed-batch mode of fermentation resulted in maximum LA titers of 97.8, 52.4 and 61.3 g/L with a yield of 0.77, 0.66 and 0.71 g/g using pure xylose, xylose-rich SCB and OP hydrolysates, respectively. Further, a two-step aqueous two-phase system (ATPS) extraction technique was employed for the separation and recovery of LA accumulated on pure and crude xylose. The LA recovery was 45 – 65% in the first step and enhanced to 80–90% in the second step.The study demonstrated an efficient integrated biorefinery approach to valorising the xylose-rich stream for cost-effective LA production and recovery.Item Open Access High level xylitol production by Pichia fermentans using non-detoxified xylose-rich sugarcane bagasse and olive pits hydrolysates(Elsevier, 2021-09-22) Narisetty, Vivek; Castro, Eulogio; Durgapal, Sumit; Coulon, Frederic; Jacob, Samuel; Kumar, Dinesh; Awasthi, Mukesh Kumar; Pant, Kamal Kishore; Parameswaran, Binod; Kumar, VinodHemicellulosic sugars, the overlooked fraction of lignocellulosic residues can serve as potential and cost-effective raw material that can be exploited for xylitol production. Xylitol is a top platform chemical with applications in food and pharmaceutical industries. Sugarcane bagasse (SCB) and olive pits (OP) are the major waste streams from sugar and olive oil industries, respectively. The current study evaluated the potential of Pichia fermentans for manufacturing of xylitol from SCB and OP hydrolysates through co-fermentation strategy. The highest xylitol accumulation was noticed with a glucose and xylose ratio of 1:10 followed by feeding with xylose alone. The fed-batch cultivation using pure xylose, SCB, and OP hydrolysates, resulted in xylitol accumulation of 102.5, 86.6 and 71.9 g/L with conversion yield of 0.78, 0.75 and 0.74 g/g, respectively. The non-pathogenic behaviour and ability to accumulate high xylitol levels from agro-industrial residues demonstrates the potential of P. fermentans as microbial cell factory.Item Embargo Paradigm of integrative OMICS of microbial technology towards biorefinery prospects(Elsevier, 2024-05-11) Jacob, Samuel; Rajeswari, Gunasekaran; Rai, Awantika; Tripathy, Sushree Shweta; Gopal, Swathy; Das, Eeshita; Kumar, Vinod; Jeevan Kumar, S. P.; Aminabhavi, Tejraj M.; Garlapati, Vijay KumarClimate change, finite natural resources, and increasing population necessitate producing sustainable energy with positive economic growth. Recent advances in OMICS coupled with genome editing and synthetic biology have paved the way for the development of sustainable technologies. These techniques help identify critical genes/pathways and re-construct and redesign biological pathways to develop eco-friendly and economically viable industrial metabolites. With the help of microbial technology, biorefinery-related research is actively pursued in many countries to develop microbial strains, producing varied value-added biochemicals and biofuels. The application of multi-omics data in deciphering key genes, their manipulation, and outcomes implies the domain's potential to find new horizons in biorefineries using microbial factories to produce various biofuels and biorefinery products. This review illustrates OMICS role in developing industrial chemicals and microbial biorefineries. Besides, prospects for genome editing and synthetic biology have been elucidated.Item Open Access Recent advances in bio-based production of top platform chemical, succinic acid: an alternative to conventional chemistry(Springer Nature, 2024-05-29) Kumar, Vinod; Kumar, Pankaj; Maity, Sunil K.; Agrawal, Deepti; Narisetty, Vivek; Jacob, Samuel; Kumar, Gopalakrishnan; Bhatia, Shashi Kant; Kumar, Dinesh; Vivekanand, VivekanandSuccinic acid (SA) is one of the top platform chemicals with huge applications in diverse sectors. The presence of two carboxylic acid groups on the terminal carbon atoms makes SA a highly functional molecule that can be derivatized into a wide range of products. The biological route for SA production is a cleaner, greener, and promising technological option with huge potential to sequester the potent greenhouse gas, carbon dioxide. The recycling of renewable carbon of biomass (an indirect form of CO2), along with fixing CO2 in the form of SA, offers a carbon-negative SA manufacturing route to reduce atmospheric CO2 load. These attractive attributes compel a paradigm shift from fossil-based to microbial SA manufacturing, as evidenced by several commercial-scale bio-SA production in the last decade. The current review article scrutinizes the existing knowledge and covers SA production by the most efficient SA producers, including several bacteria and yeast strains. The review starts with the biochemistry of the major pathways accumulating SA as an end product. It discusses the SA production from a variety of pure and crude renewable sources by native as well as engineered strains with details of pathway/metabolic, evolutionary, and process engineering approaches for enhancing TYP (titer, yield, and productivity) metrics. The review is then extended to recent progress on separation technologies to recover SA from fermentation broth. Thereafter, SA derivatization opportunities via chemo-catalysis are discussed for various high-value products, which are only a few steps away. The last two sections are devoted to the current scenario of industrial production of bio-SA and associated challenges, along with the author's perspective.Item Open Access Unlocking the potential of insect and ruminant host symbionts for recycling of lignocellulosic carbon with a biorefinery approach: a review(BMC (part of Springer Nature), 2021-05-27) Rajeswari, Gunasekaran; Jacob, Samuel; Chandel, Anuj Kumar; Kumar, VinodUprising fossil fuel depletion and deterioration of ecological reserves supply have led to the search for alternative renewable and sustainable energy sources and chemicals. Although first generation biorefinery is quite successful commercially in generating bulk of biofuels globally, the food versus fuel debate has necessitated the use of non-edible feedstocks, majorly waste biomass, for second generation production of biofuels and chemicals. A diverse class of microbes and enzymes are being exploited for biofuels production for a series of treatment process, however, the conversion efficiency of wide range of lignocellulosic biomass (LCB) and consolidated way of processing remains challenging. There were lot of research efforts in the past decade to scour for potential microbial candidate. In this context, evolution has developed the gut microbiota of several insects and ruminants that are potential LCB degraders host eco-system to overcome its host nutritional constraints, where LCB processed by microbiomes pretends to be a promising candidate. Synergistic microbial symbionts could make a significant contribution towards recycling the renewable carbon from distinctly abundant recalcitrant LCB. Several studies have assessed the bioprospection of innumerable gut symbionts and their lignocellulolytic enzymes for LCB degradation. Though, some reviews exist on molecular characterization of gut microbes, but none of them has enlightened the microbial community design coupled with various LCB valorization which intensifies the microbial diversity in biofuels application. This review provides a deep insight into the significant breakthroughs attained in enrichment strategy of gut microbial community and its molecular characterization techniques which aids in understanding the holistic microbial community dynamics. Special emphasis is placed on gut microbial role in LCB depolymerization strategies to lignocellulolytic enzymes production and its functional metagenomic data mining eventually generating the sugar platform for biofuels and renewable chemicals production.