Intracellular cellobiose metabolism and its applications in lignocellulose-based biorefineries

Parisutham, Vinuselvi; Sathesh‐Prabu, Chandran; Mukhopadhyay, Aindrila; Lee, Sung Kuk; Keasling, Jay D.

doi:10.1016/j.biortech.2017.05.001

Cited by 73 publications

(51 citation statements)

References 95 publications

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“…It is a by-product of cellulose saccharification with standard commercial mixtures of cellulases but becomes a predominant product when β-glucosidase is omitted from the cocktail (Chen, 2015). Well-defined microbial hosts capable of efficient cellobiose utilisation are therefore desirable because they can be applied in simultaneous saccharification and fermentation of cellulose for production of VAC while the process cost is reduced as addition of expensive β-glucosidase is not needed (Ha et al , 2011; Chen, 2015; Parisutham et al , 2017).…”

Section: Resultsmentioning

confidence: 99%

Biotransformation of D-xylose to D-xylonic acid coupled to medium chain length polyhydroxyalkanoate production in cellobiose-grownPseudomonas putidaEM42

Dvořák

Kováč

Lorenzo

2019

Preprint

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1Co-production of two or more desirable compounds from low-cost substrates by a single 2 microbial catalyst could greatly improve the economic competitiveness of many 3 biotechnological processes. However, reports demonstrating the adoption of such co-4 production strategy are still scarce. In this study, the ability of genome-edited strain 5 Psudomonas putida EM42 to simultaneously valorise D-xylose and D-cellobiose -two 6 important lignocellulosic carbohydrates -by converting them into the platform chemical D-7 xylonic acid and medium chain length polyhydroxyalkanoates, respectively, was investigated. 8 Biotransformation experiments performed with P. putida resting cells showed that 9 promiscuous periplasmic glucose oxidation route can efficiently generate extracellular 10 xylonate with high yield reaching 0.97 g per g of supplied xylose. Xylose oxidation was 11 subsequently coupled to the growth of P. putida with cytoplasmic -glucosidase BglC from 12 Thermobifida fusca on D-cellobiose. This disaccharide turned out to be a better co-substrate 13 for xylose-to-xylonate biotransformation than monomeric glucose. This was because unlike 14 glucose, cellobiose did not block oxidation of the pentose by periplasmic glucose 15 dehydrogenase Gcd, but, similarly to glucose, it was a suitable substrate for 16 polyhydroxyalkanoate formation in P. putida. Co-production of extracellular xylose-born 17 xylonate and intracellular cellobiose-born medium chain length polyhydroxyalkanoates was 18 established in proof-of-concept experiments with P. putida grown on the disaccharide. This 19 study highlights the potential of P. putida EM42 as a microbial platform for the production of 20 xylonic acid, identifies cellobiose as a new substrate for mcl-PHA production, and proposes a 21 fresh strategy for the simultaneous valorisation of xylose and cellobiose.22 23 24 3

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Section: Resultsmentioning

confidence: 99%

Biotransformation of D-xylose to D-xylonic acid coupled to medium chain length polyhydroxyalkanoate production in cellobiose-grownPseudomonas putidaEM42

Dvořák

Kováč

Lorenzo

2019

Preprint

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“…Alternative strategies included intracellular assimilation of cellobiose via endogenous or exogenous β-glucosidase (Ha et al, 2011; Vinuselvi and Lee, 2011), and via phosphorolysis catalyzed by cellobiose phosphorylase (Shin et al, 2014). Despite the need for an efficient cellobiose transporter, intracellular assimilation is generally regarded as more efficient, because it circumvents carbon catabolite repression (CCR) and prevents cellobiose inhibition of extracellular cellulases in the bioreactor (Ha et al, 2011; Parisutham et al, 2017; Teugjas and Väljamäe, 2013).…”

Section: Resultsmentioning

confidence: 99%

“…Uptake of cellobiose and other cellodextrins of varying lengths through the ABC-type transporters is common in cellulolytic bacteria due to the relatively broad specificity of these systems (Nataf et al, 2009; Parisutham et al, 2017). To test its relevance for cellobiose uptake in EM42 strain, we deleted the gtsABCD operon that encodes the ABC glucose transporter.…”

Section: Resultsmentioning

confidence: 99%

“…In the case, for instance, of E. coli with its characteristic Embden– Meyerhof–Parnas pathway, the ATP yield per molecule of glucose is twice as high. It is thought that environmental or engineered microorganisms that prefer to metabolize cellobiose instead of glucose are more energetic and robust than their glucose-utilizing counterparts (Chen, 2015; Lynd et al, 2002; Parisutham et al, 2017). The benefits were demonstrated in bacteria with a cellobiose-specific PEP-phosphotransferase system transporter, in which one mole of ATP is consumed per one mole of imported substrate; this was also apparent in microbes that metabolize cellobiose through phosphorolysis, in which only one ATP per disaccharide is needed to form two activated molecules of glucose (Kajikawa and Masaki, 1999; Shin et al, 2014; Thurston et al, 1993).…”

Section: Resultsmentioning

confidence: 99%

“…Cellodextrins, including D-cellobiose, are the most abundant by-products of cellulose saccharification and predominate following partial hydrolysis (Chen, 2015; Singhania et al, 2013). Well defined, industrially relevant microbes with efficient cellobiose metabolism are thus very desirable (Kawaguchi et al, 2016; Parisutham et al, 2017; Taha et al, 2016). Both cellobiose and xylose metabolism have been established artificially in a number of microorganisms (Ha et al, 2011; Lane et al, 2015; Le Meur et al, 2012; Lee et al, 2016; Meijnen et al, 2008; Shin et al, 2014; Vinuselvi and Lee, 2011) and some Pseudomonas species can use xylose as a C source (Liu et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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Refactoring the upper sugar metabolism ofPseudomonas putidafor co-utilization of disaccharides, pentoses, and hexoses

Dvořák

Lorenzo

2018

Preprint

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6Running title: Engineering cellobiose and xylose metabolism in P. putida 7 Abstract 1 Given its capacity to tolerate stress, NAD(P)H/ NAD(P) balance, and increased ATP levels, 2 the platform strain Pseudomonas putida EM42, a genome-edited derivative of the soil 3 bacterium P. putida KT2440, can efficiently host a suite of harsh reactions of biotechnological 4 interest. Because of the lifestyle of the original isolate, however, the nutritional repertoire of P. 5 putida EM42 is centered largely on organic acids, aromatic compounds and some hexoses 6 (glucose and fructose). To enlarge the biochemical network of P. putida EM42 to include 7 disaccharides and pentoses, we implanted heterologous genetic modules for D-cellobiose and 8 D-xylose metabolism into the enzymatic complement of this strain. Cellobiose was actively 9 transported into the cells through the ABC complex formed by native proteins PP1015-PP1018. 10The knocked-in -glucosidase BglC from Thermobifida fusca catalyzed intracellular cleavage 11 of the disaccharide to D-glucose, which was then channelled to the default central metabolism. 12Xylose oxidation to the dead end product D-xylonate was prevented by by deleting the gcd 13 gene that encodes the broad substrate range quinone-dependent glucose dehydrogenase. 14 Intracellular intake was then engineered by expressing the Escherichia coli proton-coupled 15 symporter XylE. The sugar was further metabolized by the products of E. coli xylA (xylose 16 isomerase) and xylB (xylulokinase) towards the pentose phosphate pathway. The resulting P. 17 putida strain co-utilized xylose with glucose or cellobiose to complete depletion of the sugars. 18 These results not only show the broadening of the metabolic capacity of a soil bacterium 19 towards new substrates, but also promote P. putida EM42 as a platform for plug-in of new 20 biochemical pathways for utilization and valorization of carbohydrate mixtures from 21 lignocellulose processing.22 23 24recruitment of one heterologous -glucosidase for intracellular cellobiose hydrolysis, and the 1 implementation of three enterobacterial genes (xylose transporter, isomerase and kinase) 2 sufficed to cause disaccharide and the pentose co-utilization by P. putida EM42 with 3 inactivated glucose dehydrogenase while maintaining its ability to use glucose. We also 4 demonstrate that P. putida metabolism generates more ATP when cells are grown on cellobiose 5 instead of glucose. This study expands the catalytic scope of P. putida towards utilization of 6 major components of all three lignocellulose-derived fractions. Moreover, given that the 7 cellobiose, as the bulk by-product of standard cellulose saccharification, frequently remains 8 untouched in the sugar mix (due to the inability of most microorganisms to assimilate it), our 9 results demonstrate rational tailoring of an industrially relevant microbial host to achieve a 10 specific step in such a biotechnological value chain. 11 12 13 14 6 1 Figure 1. Engineering Pseudomonas putida EM42 for co-utilization of D-cellobios...

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Current challenges of biomass refinery and prospects of emerging technologies for sustainable bioproducts and bioeconomy

Asghar

Sairash

Hussain

et al. 2022

Biofuels Bioprod Bioref

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The depletion of fossil resources and the resulting global warming are the primary motivators for moving from a fossil‐based to a biobased economy. Biorefineries, like petro‐based refineries, offer a green and sustainable option for producing marketable biobased goods through decarbonization pathways, hence upgrading the bioeconomy. In the context of the industrial revolution, economic and environmental sustainability are critical in adopting biorefinery systems. However, biorefineries still face numerous hurdles. There are numerous technological, economic, ecological, sociological and long‐term issues throughout the biorefinery production chain. The key difficulties facing biorefineries today are acceptance in the present market of the ‘fossil‐based economy’, the composition and availability of feedstock, the quantities needed to meet the market demand, the efficiency of the resource recovery, techno‐economic viability and sustainability. Life cycle assessment may provide a thorough input on the environmental setting during process optimization, minimizing the biorefinery project's environmental impact. In this review, we outline the bottlenecks and obstacles that biorefineries face while producing biofuels from biomass, as well as how biotechnology and modern technology might help researchers to overcome them. Biorefineries need to optimize their biomass usage to generate products that properly fit the market expectations. These items should be cost‐effective when compared with fossil fuels. Currently, 85–90% of petroleum refinery output is used to make fuels, with only 10–15% going to the petrochemical industry to make organic compounds. To precisely match market demands, the biorefinery should produce the same proportion of fuels and organic compounds. To overcome the challenges, biofermentation, membrane technologies and oxidation processes, among other developing technologies, are drawing interest in several areas of biorefinery. Biorefinery systems necessitate a systematic process for identifying impacts and assessing their long‐term viability. © 2022 Society of Chemical Industry and John Wiley & Sons, Ltd.

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Intracellular cellobiose metabolism and its applications in lignocellulose-based biorefineries

Cited by 73 publications

References 95 publications

Biotransformation of D-xylose to D-xylonic acid coupled to medium chain length polyhydroxyalkanoate production in cellobiose-grownPseudomonas putidaEM42

Biotransformation of D-xylose to D-xylonic acid coupled to medium chain length polyhydroxyalkanoate production in cellobiose-grownPseudomonas putidaEM42

Refactoring the upper sugar metabolism ofPseudomonas putidafor co-utilization of disaccharides, pentoses, and hexoses

Current challenges of biomass refinery and prospects of emerging technologies for sustainable bioproducts and bioeconomy

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