Bioconversion of plant biomass hydrolysate into bioplastic (polyhydroxyalkanoates) using <i>Ralstonia eutropha</i> 5119

Bhatia, Shashi Kant,Gurav, Ranjit,Choi, Tae-Rim,Jung, Hye-Rim,Yang, Soo-Yeon,Moon, Yu-Mi,Song, Hun-Suk,Jeon, Jong-Min,Choi, Kwon-Young,Yang, Yung-Hun Elsevier 2019 Bioresource technology Vol.271 No.-

<P><B>Abstract</B></P> <P>Pretreatment of lignocellulosic biomass results in the formation of byproducts (furfural, hydroxymethylfurfural [HMF], vanillin, acetate etc.), which affect microbial growth and productivity. Furfural (0.02%), HMF (0.04%), and acetate (0.6%) showed positive effects on <I>Ralstonia eutropha</I> 5119 growth and polyhydroxyalkanoate (PHA) production, while vanillin exhibited negative effects. Response optimization and interaction studies between the variables glucose, ammonium chloride, furfural, HMF, and acetate using the response surface methodology resulted in maximum PHA production (2.1 g/L) at optimal variable values of 15.3 g/L, 0.43 g/L, 0.04 g/L, 0.05 g/L, and 2.34 g/L, respectively. Different lignocellulosic biomass hydrolysates (LBHs), including barley biomass hydrolysate (BBH), <I>Miscanthus</I> biomass hydrolysate (MBH), and pine biomass hydrolysate (PBH), were evaluated as potential carbon sources for <I>R. eutropha</I> 5119 and resulted in 1.8, 2.0, and 1.7 g/L PHA production, respectively. MBH proved the best carbon source, resulted in higher biomass (Y<SUB>x/s,</SUB> 0.31 g/g) and PHA (Y<SUB>p/s,</SUB> 0.14 g/g) yield.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <I>Ralstonia eutropha</I> 5119 can co-metabolize biomass derived byproducts with glucose. </LI> <LI> Furfural, hydroxymethylfurfural and acetate promote biomass and PHA production. </LI> <LI> Vanillin is more toxic followed by furfural > hydroxymethylfurfural > acetate. </LI> <LI> <I>Miscanthus</I> biomass hydrolysate resulted in high PHA (Y<SUB>p/s,</SUB> 0.14 g/g) yield. </LI> <LI> PHA produced from biomass hydrolysate has similar properties to P(3HB-<I>co</I>-3HV). </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) production from engineered Ralstonia eutropha using synthetic and anaerobically digested food waste derived volatile fatty acids

Bhatia, Shashi Kant,Gurav, Ranjit,Choi, Tae-Rim,Jung, Hye-Rim,Yang, Soo-Yeon,Song, Hun-Suk,Jeon, Jong-Min,Kim, Jae-Seok,Lee, Yoo-Kyung,Yang, Yung-Hun Elsevier 2019 INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES Vol.133 No.3

Engineering of artificial microbial consortia of <i>Ralstonia eutropha</i> and <i>Bacillus subtilis</i> for poly(3-hydroxybutyrate-<i>co</i>-3-hydroxyvalerate) copolymer production from sugarcane sugar without precursor feeding

Bhatia, Shashi Kant,Yoon, Jeong-Jun,Kim, Hyun-Joong,Hong, Ju Won,Gi Hong, Yoon,Song, Hun-Seok,Moon, Yu-Mi,Jeon, Jong-Min,Kim, Yun-Gon,Yang, Yung-Hun Elsevier 2018 Bioresource technology Vol.257 No.-

<P><B>Abstract</B></P> <P> <I>Ralstonia eutropha</I> is a well-known microbe reported for polyhydroxyalkonate (PHA) production, and unable to utilize sucrose as carbon source. Two strains, <I>Ralstonia eutropha</I> H16 and <I>Ralstonia eutropha</I> 5119 were co-cultured with sucrose hydrolyzing microbes (<I>Bacillus subtilis</I> and <I>Bacillus amyloliquefaciens</I>) for PHA production<I>.</I> Co-culture of <I>B. subtilis:R. eutropha</I> 5119 (BS:RE5) resulted in best PHA production (45% w/w dcw). Optimization of the PHA production process components through response surface resulted in sucrose: NH<SUB>4</SUB>Cl:<I>B. subtilis</I>: <I>R. eutropha</I> (3.0:0.17:0.10:0.190). Along with the hydrolysis of sucrose, <I>B. subtilis</I> also ferments sugars into organic acid (propionic acid), which acts as a precursor for HV monomer unit. Microbial consortia of BS:RE5 when cultured in optimized media led to the production of poly(3-hydroxybutyrate-<I>co</I>-3-hydroxyvalerate) (P(3HB-<I>co</I>-3HV) with 66% w/w of dcw having 16 mol% HV fraction<I>.</I> This co-culture strategy overcomes the need for metabolic engineering of <I>R. eutropha</I> for sucrose utilization, and addition of precursor for copolymer production.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <I>Ralstonia eutropha</I> 5119 strain unable to utilize sucrose as carbon source. </LI> <LI> <I>Bacillus subtilis</I> hydrolyze sucrose into free sugars and produce propionic acid. </LI> <LI> <I>Ralstonia eutropha</I> 5119: <I>Bacillus subtilis</I> produce P(3HB-<I>co</I>-3HV) by 66% w/w of dcw. </LI> <LI> Population dynamics study shows both microbes are compatible with each other. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Effect of synthetic and food waste-derived volatile fatty acids on lipid accumulation in <i>Rhodococcus</i> sp. YHY01 and the properties of produced biodiesel

Bhatia, Shashi Kant,Gurav, Ranjit,Choi, Tae-Rim,Jung, Hye-Rim,Yang, Soo-Yeon,Song, Hun-Suk,Kim, Yun-Gon,Yoon, Jeong-Jun,Yang, Yung-Hun Elsevier 2019 Energy conversion and management Vol.192 No.-

<P><B>Abstract</B></P> <P>Food waste-derived volatile fatty acids (VFAs) can act as a renewable feedstock for biodiesel production. In synthetic media, <I>Rhodococcus</I> sp. YHY01 was able to utilize various organic acids (acetate, butyrate, lactate, and propionate) as a carbon source. Butyrate was the optimal carbon source, having a minimum inhibitory effect on growth, and a maximum growth yield coefficient (Y<SUB>x/s</SUB> 0.288 g dcw/g butyrate) and fatty acid yield coefficient (Y<SUB>f/s</SUB> 0.206 g/g butyrate), compared to other organic acids (lactate, propionate, and acetate). Acetate, butyrate, and lactate mostly supported the production of fatty acids with an even number of carbons, whereas propionate enhanced the content of odd-numbered fatty acids. Response surface methodology (RSM) design study resulted in maximum biomass (2.8 g/L) and fatty acid yield (1.9 g/g) with acetate:butyrate:lactate (0.333:0.333:0.333) as a carbon source. Culture of <I>Rhodococcus</I> sp. YHY01 in media containing food waste-derived VFAs as the carbon source had a biomass (3.2 g dcw/L), fatty acid yield (2.2 g/L), and fatty acid accumulation (69% w/w) under nitrogen-limited condition. Biodiesel produced from food waste had an iodine value (IV, 37), cetane number (CN, 63), high heating value (HHV, 39), density (υ, 3.9), and viscosity (ρ, 0.868) that meet international standards.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <I>Rhodococcus</I> sp. YHY01 can utilize volatile fatty acids as carbon source. </LI> <LI> Acetate, butyrate and lactate play role in even number fatty acids synthesis. </LI> <LI> Propionate directly involved in synthesis of odd carbon number fatty acids. </LI> <LI> Higher biomass and fatty acid yield coefficient obtained with butyrate. </LI> <LI> Food waste derived volatile fatty acids are a suitable feedstock for biodiesel production. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Bioconversion of barley straw lignin into biodiesel using <i>Rhodococcus</i> sp. YHY01

Bhatia, Shashi Kant,Gurav, Ranjit,Choi, Tae-Rim,Han, Yeong Hoon,Park, Ye-Lim,Park, Jun Young,Jung, Hye-Rim,Yang, Soo-Yeon,Song, Hun-Suk,Kim, Sang-Hyoun,Choi, Kwon-Young,Yang, Yung-Hun Elsevier 2019 Bioresource technology Vol.289 No.-

<P><B>Abstract</B></P> <P> <I>Rhodococcus</I> sp. YHY01 was studied to utilize various lignin derived aromatic compounds. It was able to utilize <I>p</I>-coumaric acid, cresol, and 2,6 dimethoxyphenol and resulted in biomass production i.e. 0.38 g dcw/L, 0.25 g dcw/L and 0.1 g dcw/L, and lipid accumulation i.e. 49%, 40%, 30%, respectively. The half maximal inhibitory concentration (IC<SUB>50</SUB>) value for <I>p</I>-coumaric acid (13.4 mM), cresol (7.9 mM), and 2,6 dimethoxyphenol (3.4 mM) was analyzed. Dimethyl sulfoxide (DMSO) solubilized barley straw lignin fraction was used as a carbon source for <I>Rhodococcus</I> sp. YHY01 and resulted in 0.130 g dcw/L with 39% w/w lipid accumulation. Major fatty acids were palmitic acid (C16:0) 51.87%, palmitoleic acid (C16:l) 14.90%, and oleic acid (C18:1) 13.76%, respectively. Properties of biodiesel produced from barley straw lignin were as iodine value (IV) 27.25, cetane number (CN) 65.57, cold filter plugging point (CFPP) 14.36, viscosity (υ) 3.81, and density (ρ) 0.86.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <I>Rhodococcus</I> sp. YHY01 can utilize <I>p</I>-coumaric acid > cresol > 2,6 dimethoxyphenol. </LI> <LI> IC<SUB>50</SUB> value was; <I>p</I>-coumaric (13.4 mM), cresol (7.9 mM), 2,6 dimethoxyphenol (3.4 mM) </LI> <LI> Biomass and lipid production in order; <I>p</I>-coumaric acid > cresol > 2,6 dimethoxyphenol. </LI> <LI> Barley lignin led to 39% w/w lipid accumulation in <I>Rhodococcus</I> sp. YHY01. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Lipase mediated functionalization of poly(3-hydroxybutyrate-<i>co</i>-3-hydroxyvalerate) with ascorbic acid into an antioxidant active biomaterial

Bhatia, Shashi Kant,Wadhwa, Puneet,Hong, Ju Won,Hong, Yoon Gi,Jeon, Jong-Min,Lee, Eui Seok,Yang, Yung-Hun Elsevier 2019 INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES Vol.123 No.-

<P><B>Abstract</B></P> <P>Naturally produced polyhydroxyalkanoates (PHAs) biopolymers have limited medical applications due to their brittle and hydrophobic nature. In this study poly(3-hydroxybutyrate-<I>co</I>-3-hydroxyvalerate) P(3HB-<I>co</I>-3HV) copolymer was produced using engineered <I>Escherichia coli</I> YJ101, and further functionalized with ascorbic acid using <I>Candida antarctica</I> lipase B mediated esterification. Copolymer P(3HB-<I>co</I>-3HV)-ascorbic acid showed lower degree of crystallinity (9.96%), higher thermal degradation temperature (294.97 °C) and hydrophilicity (68°) as compared to P(3HB-<I>co</I>-3HV). Further, P(3HB-<I>co</I>-3HV)-ascorbic acid biomaterial showed 14% scavenging effect on 1,1-diphenyl-2-picryl-hydrazyl (DPPH), and 1.6 fold increase in biodegradability as compared to P(3HB-<I>co</I>-3HV). Improvement of PHAs polymer properties by adding functional groups could be a good approach to increase their biodegradability, economic value and important applications in the medical field.</P>

Microbial biodiesel production from oil palm biomass hydrolysate using marine <i>Rhodococcus</i> sp. YHY01

Bhatia, Shashi Kant,Kim, Junyoung,Song, Hun-Seok,Kim, Hyun Joong,Jeon, Jong-Min,Sathiyanarayanan, Ganesan,Yoon, Jeong-Jun,Park, Kyungmoon,Kim, Yun-Gon,Yang, Yung-Hun Elsevier Applied Science 2017 Bioresource technology Vol.233 No.-

<P><B>Abstract</B></P> <P>The effect of various biomass derived inhibitors (i.e. furfural, hydroxymethylfurfural (HMF), vanillin, 4-hydroxy benzaldehyde (4-HB) and acetate) was investigated for fatty acid accumulation in <I>Rhodococcus</I> sp. YHY 01. <I>Rhodococcus</I> sp. YHY01 was able to utilize acetate, vanillin, and 4-HB for biomass production and fatty acid accumulation. The IC<SUB>50</SUB> value for furfural (3.1mM), HMF (3.2mM), vanillin (2.0mM), 4-HB (2.7mM) and acetate (3.7mM) was calculated. HMF and vanillin affect fatty acid composition and increase saturated fatty acid content. <I>Rhodococcus</I> sp. YHY 01 cultured with empty fruit bunch hydrolysate (EFBH) as the main carbon source resulted in enhanced biomass (20%) and fatty acid productivity (37%), in compression to glucose as a carbon source. Overall, this study showed the beneficial effects of inhibitory molecules on growth and fatty acid production, and support the idea of biomass hydrolysate utilization for biodiesel production by avoiding complex efforts to remove inhibitory compounds.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <I>Rhodococcus</I> sp. YHY 01 is able to utilize acetate, 4-HB and vanillin as carbon source. </LI> <LI> HMF, 4-HB and vanillin enhance saturated fatty acid content. </LI> <LI> Salt stress increases the content of unsaturated fatty acids. </LI> <LI> Production in EFBH hydrolysate results in 69% fatty acid accumulation. </LI> <LI> EFBH hydrolysate enhanced fatty acid accumulation by 37% in comparison to glucose. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

An overview of microdiesel — A sustainable future source of renewable energy

Bhatia, Shashi Kant,Bhatia, Ravi Kant,Yang, Yung-Hun Elsevier 2017 RENEWABLE & SUSTAINABLE ENERGY REVIEWS Vol.79 No.-

<P><B>Abstract</B></P> <P>Microdiesel obtained from microbes using renewable materials as carbon sources is an important alternative to petroleum diesel. This review provides information related to microdiesel production using various carbon sources; i.e. carbon dioxide, C<SUB>2</SUB>, saccharides, and lignocellulose. Microbes can accumulate different contents of fatty acids in the form of triacylglycerol (TAG). Not all microbes store fatty acids and utilize a broad range of substrates as carbon sources, and vice versa. Microbes can be engineered to consume various carbon sources, and accumulate increased amounts of fatty acids with different composition. The properties of microdiesel depend on its fatty acid profile, which in turn determines its efficacy. The structural features of the fatty acids, such as carbon chain length, branching and degree of unsaturation, affect the physiochemical properties of the biodiesel (cetane number (CN), oxidation stability (OS), iodine value (IV), cold flow properties, density and kinematic viscosity). Fatty acid methyl ester (FAME) profiles can be used to evaluate the key properties of biodiesel, i.e. the stability of the oil used. The overview presented herein concludes that microdiesel production using non-feed carbon sources and genetically engineered microbes shows much promise.</P>

Starch based polyhydroxybutyrate production in engineered Escherichia coli.

Bhatia, Shashi Kant,Shim, Young-Ha,Jeon, Jong-Min,Brigham, Christopher J,Kim, Yong-Hyun,Kim, Hyun-Joong,Seo, Hyung-Min,Lee, Ju-Hee,Kim, Jung-Ho,Yi, Da-Hye,Lee, Yoo Kyung,Yang, Yung-Hun Springer-Verlag 2015 Bioprocess and biosystems engineering Vol.38 No.8

<P>Every year, the amount of chemosynthetic plastic accumulating in the environment is increasing, and significant time is required for decomposition. Bio-based, biodegradable plastic is a promising alternative, but its production is not yet a cost effective process. Decreasing the production cost of polyhydroxyalkanoate by utilizing renewable carbon sources for biosynthesis is an important aspect of commercializing this biodegradable polymer. An Escherichia coli strain that expresses a functional amylase and accumulate polyhydroxybutyrate (PHB), was constructed using different plasmids containing the amylase gene of Panibacillus sp. and PHB synthesis genes from Ralstonia eutropha. This engineered strain can utilize starch as the sole carbon source. The maximum PHB production (1.24?g/L) was obtained with 2?% (w/v) starch in M9 media containing 0.15?% (w/v) yeast extract and 10?mM glycine betaine. The engineered E. coli SKB99 strain can accumulate intracellular PHB up to 57.4?% of cell dry mass.</P>

Biowaste-to-bioenergy using biological methods – A mini-review

Bhatia, Shashi Kant,Joo, Hwang-Soo,Yang, Yung-Hun Elsevier 2018 Energy conversion and management Vol.177 No.-

<P><B>Abstract</B></P> <P>The continued production of waste is creating management problems. The use of traditional waste management methods, such as incineration and landfill, releases gases that may cause global warming. Energy demand is also increasing rapidly owing to the rapid increase in population and industrialization. To meet this ever-increasing demand, access to clean and green energy is essential for the sustainable development of human society. These two challenges, if managed scientifically using biowaste to bioenergy (BtB) technology, can provide solutions for one another. In this article, we reviewed the strategies for and status of BtB technology (anaerobic digestion, transesterification, and microbial fuel cells) used to convert various biowastes (forest and agriculture residue, animal wastes, and municipal wastes) into bioenergy (biogas, biodiesel, bioalcohol, and bioelectricity). The participation of researchers, scientists, government agencies, and stakeholders is needed to increase the feasibility of these technologies.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Biowaste-to-bioenergy technology is a possible solution to fulfill energy demand. </LI> <LI> This technology will not only solve energy problem but also help to manage biowaste. </LI> <LI> There is need to develop an integrated process to get more revenue from biowaste. </LI> <LI> To compete with other energy source this technology need government policy and subsidies. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>