Mary Grace Baylon

High-level conversion of L-lysine into 5-aminovalerate that can be used for nylon 6,5 synthesis

L-Lysine is a potential feedstock for the production of bio-based precursors for engineering plastics. In this study, we developed a microbial process for high-level conversion of L-lysine into 5-aminovalerate (5AVA) that can be used as a monomer in nylon 6,5 synthesis. Recombinant Escherichia coli WL3110 strain expressing Pseudomonas putida delta-aminovaleramidase (DavA) and lysine 2-monooxygenase (DavB) was grown to high density in fed-batch culture and used as a whole cell catalyst. High-density E. coli WL3110 expressing DavAB, grown to an optical density at 600 nm (OD600 ) of 30, yielded 36.51 g/L 5AVA from 60 g/L L-lysine in 24 h. Doubling the cell density of E. coli WL3110 improved the conversion yield to 47.96 g/L 5AVA from 60 g/L of L-lysine in 24 h. 5AVA production was further improved by doubling the L-lysine concentration from 60 to 120 g/L. The highest 5AVA titer (90.59 g/L; molar yield 0.942) was obtained from 120 g/L L-lysine by E. coli WL3110 cells grown to OD600 of 60. Finally, nylon 6,5 was synthesized by bulk polymerization of ϵ-caprolactam and δ-valerolactam prepared from microbially synthesized 5AVA. The hybrid system demonstrated here has promising possibilities for application in the development of industrial bio-nylon production processes.

Metabolic Engineering of Ralstonia eutropha for the Production of Polyhydroxyalkanoates From Sucrose

A sucrose utilization pathway was established in Ralstonia eutropha NCIMB11599 and R. eutropha 437-540 by introducing the Mannheimia succiniciproducens MBEL55E sacC gene that encodes β-fructofuranosidase. These engineered strains were examined for the production of poly(3-hydroxybutyrate) [P(3HB)] and poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)], respectively, from sucrose as a carbon source. It was found that β-fructofuranosidase excreted into the culture medium could hydrolyze sucrose to glucose and fructose, which were efficiently used as carbon sources by recombinant R. eutropha strains. When R. eutropha NCIMB11599 expressing the sacC gene was cultured in nitrogen-free chemically defined medium containing 20 g/L of sucrose, a high P(3HB) content of 73.2 wt% could be obtained. In addition, R. eutropha 437-540 expressing the Pseudomonas sp. MBEL 6-19 phaC1437 gene and the Clostridium propionicum pct540 gene accumulated P(3HB-co-21.5 mol% LA) to a polymer content of 19.5 wt% from sucrose by the expression of the sacC gene and the Escherichia coli ldhA gene. The molecular weights of P(3HB) and P(3HB-co-21.5 mol%LA) synthesized in R. eutropha using sucrose as a carbon source were 3.52 × 10(5) (Mn ) and 2.19 × 10(4) (Mn ), respectively. The engineered R. eutropha strains reported here will be useful for the production of polyhydroxyalkanoates (PHAs) from sucrose, one of the most abundant and relatively inexpensive carbon sources.

Development of rice bran treatment process and its use for the synthesis of polyhydroxyalkanoates from rice bran hydrolysate solution

Rice bran treatment process for the production of 43.7 kg of hydrolysate solution containing 24.41 g/L of glucose and small amount of fructose from 5 kg of rice bran was developed and employed to produce polyhydroxyalkanoates in recombinant Escherichia coli and Ralstonia eutropha strains. Recombinant E. coli XL1-Blue expressing R. eutropha phaCAB genes and R. eutropha NCIMB11599 could produce poly(3-hydroxybutyrate) with the polymer contents of 90.1 wt% and 97.2 wt%, respectively, when they were cultured in chemically defined MR medium and chemically defined nitrogen free MR medium containing 10 mL/L of rice bran hydrolysate solution, respectively. Also, recombinant E. coli XL1-Blue and recombinant R. eutropha 437-540, both of which express the Pseudomonas sp. phaC1437 gene and the Clostridium propionicum pct540 gene could produce poly(3-hydroxybutyrate-co-lactate) from rice bran hydrolysate solution. These results suggest that rice bran may be a good renewable resource for the production of biomass-based polymers by recombinant microorganisms.

Advances in the biological treatment of coal for synthetic natural gas and chemicals

Coal, the most primitive fossil fuel, has been exploited for ages, and reserves dictate the economies of many countries. Presently, most energy is generated by direct combustion, raising concerns over global warming. Biological pretreatment of fossil resources and generation of alternative green energy can address the environmental issues associated with global coal utilization. Biological coal treatment can produce industrially important chemicals and bio-methane by employing microorganisms able to depolymerize/degrade coal. This review discusses current advances in microbial coal conversion, such as the efforts made to comprehend microbial processes, significant outputs of coal conversion, principle components responsible for coal conversion, and factors affecting the biological processes to convert coal. Development of these biological processes can be a stepping stone for greener coal; however, integration of multidisciplinary technologies is needed to increase the efficiency of economic coal utilization and production of economically and industrially feasible biomethane.

Bio-solubilization of the untreated low rank coal by alkali-producing bacteria isolated from soil

Coal is a hydrocarbon-rich fossil fuel considered as a possible replacement for petroleum as a feedstock for the production of fuel and valuable chemicals. In this study, bacteria capable of solubilizing untreated low rank coal were isolated from soil. A total of 19 microorganisms were isolated from soil enriched in MR medium with coal and were identified based on 16S rRNA sequencing. The identified soil isolates belonging to the genera Citricoccus, Comamonas, Cupriavidus, Sphingomonas, and Sphingopyxis were screened based on their growth in the chemically defined MR medium containing different concentrations of coal. Among the identified microbial strains, Cupriavidus necator S2A2, Sphingopyxis ginsengisoli S2B14 and Sphingomonas sp. S2B18 were further characterized for their ability to degrade low-rank coal. Cupriavidus necator S2A2, Sphingopyxis ginsengisoli S2B14 and Sphingomonas sp. S2B18 were found to solubilize untreated low-rank coal as indicated by the release of solubilized coal products detected at OD450 when they were grown in LB medium containing 1% coal. Sphingomonas sp. S2B18 showed the highest coal solubilization activity, based on the high absorbance of its culture supernatant (0.190). Although laccase-like activity was not detected in these strains when tested for RBBR dye degradation, increase in the pH of the culture medium up to 8.25- 8.34 was observed. This may be attributed to the excretion of alkaline substances in the culture medium. Since biosolubilization of coal by microorganisms is a good alternative for the chemical conversion of coal, microorganisms screened in this study can be potentially used as biological catalysts for the conversion of coal into valuable chemicals.