Chitin and Chitosan: Sustainable, Medically Relevant Biomaterials

The polysaccharides chitin and chitosan are made up of monomer units of the amino sugars D-glucosamine and N-acetyl-D-glucosamine. The ratio of these two monomers dictates whether the polysaccharide is considered chitin or chitosan. Both polymers have unique properties and have uses in several diverse applications. In nature, chitin and chitosan primarily play a structural role. When purified from their producing organism, these polymers exhibit useful structural, chemical and biological properties. Chitin and chitosan have been used in several applications including biomedicine, food additives, cosmetics, and more. The charged chitosan polymer is especially effective in biomedical applications, as it has been demonstrated to possess antimicrobial properties. This review explores the properties of chitin and chitosan and how these biopolymers are used in a variety of healthcare and other applications

From Beverages to Biofuels: The Journeys of Ethanol-Producing Microorganisms

Microbial fermentation for bio-based products is quickly becoming an integral component of the world infrastructure, as the processes encompassing the synthesis of these natural products becomes more efficient and cost effective to compete with existing commodities. Bioethanol is currently one of the most desired fermentation products, as this constituent can be applied to multiple uses in not only contributing to the more traditional routes of beer brewing and winemaking, but also in the foundation for green fuel sources. By optimizing yields, the innovative processes could be applied towards engineering more rapid and productive biomanufacturing. In order to achieve these goals, we as researchers must understand the underlying principles and intricate networks that play a role within the microenvironment and also on the cellular level in key fermentative microbes such as Saccharomyces cerevisiae and Zymomonas mobilis. In-depth pathway analysis could lead to the development of more favorable metabolic outcomes. This review focuses on the key metabolic networks and cellular frameworks in these model organisms, and how biosynthesis of ethanol yields can be optimized throughout the fermentation process

Applications of Polyhydroxyalkanoates in the Medical Industry

The bio-based, biodegradable family of polymers, polyhydroxyalkanoates (PHA), is an attractive candidate for an environmentally friendly replacement of petroleum-based plastics in many applications. In the past decade, many groups have examined the biodegradability and biocompatibility of PHA in cell culture systems or in an animal host. Findings suggest that PHA is a suitable material for fabrication of resorbable medical devices, such as sutures, meshes, implants, and tissue engineering scaffolds. The degradation kinetics of some PHA polymers is also suggestive of drug release applications. In this review, we examine the progress, potential applications, challenges and outlook in the medical polyhydroxyalkanoate field

The Potential of Polyhydroxyalkanoate Production from Food Wastes

Background and objective: Over 1 billion tons of foods are wasted every year (not consumed by humans or animals). Most of this waste ends up in landfills. As the global population increases, mankind must look for more sustainable means of living. A recently popular idea is the use of organic wastes as carbon feedstocks for fermentation that produces value added products. Polyhydroxyalkanoates are a family of bio-based, biodegradable polymers that can be produced in large quantities using food and food processing wastes as the main feedstocks. In many cases, biocatalysts have been engineered to efficiently use these waste compounds to produce large quantities of useful intracellular polyhydroxyalkanoates.Results and conclusion: In the current study, various polyhydroxyalkanoates were produced; each with different thermal and mechanical characteristics useful for different applications. If polyhydroxyalkanoate production facilities are established next to food waste accumulation sites (e.g., large landfills), potentials for the economical and sustainable polyhydroxyalkanoate production sound promising.Conflict of interest: The authors declare no conflict of interest

Biosynthesis and characterization of polyhydroxyalkanoate containing high 3-hydroxyhexanoate monomer fraction from crude palm kernel oil by recombinant Cupriavidus necator

The potential of plant oils as sole carbon sources for production of P(3HB-co-3HHx) copolymer containing a high 3HHx monomer fraction using the recombinant Cupriavidus necator strain Re2160/pCB113 has been investigated. Various types and concentrations of plant oils were evaluated for efficient conversion of P(3HB-co-3HHx) copolymer. Crude palm kernel oil (CPKO) at a concentration of 2.5 g/L was found to be most suitable for production of copolymer with a 3HHx content of approximately 70 mol%. The time profile of these cells was also examined in order to study the trend of 3HHx monomer incorporation, PHA production and PHA synthase activity. [superscript 1]H NMR and [superscript 13]C NMR analyses confirmed the presence of P(3HB-co-3HHx) copolymer containing a high 3HHx monomer fraction, in which monomers were not randomly distributed. The results of various characterization analyses revealed that the copolymers containing a high 3HHx monomer fraction demonstrated soft and flexible mechanical properties.Malaysia. Ministry of Science, Technology and Innovation (MOSTI Techno Fund)Universiti Sains Malaysia (USM Fellowship

Engineered Corynebacterium glutamicum as an endotoxin-free platform strain for lactate-based polyester production

The first biosynthetic system for lactate (LA)-based polyesters was previously created in recombinant Escherichia coli (Taguchi et al. 2008). Here, we have begun efforts to upgrade the prototype polymer production system to a practical stage by using metabolically engineered Gram-positive bacterium Corynebacterium glutamicum as an endotoxin-free platform. We designed metabolic pathways in C. glutamicum to generate monomer substrates, lactyl-CoA (LA-CoA), and 3-hydroxybutyryl-CoA (3HB-CoA), for the copolymerization catalyzed by the LA-polymerizing enzyme (LPE). LA-CoA was synthesized by D-lactate dehydrogenase and propionyl-CoA transferase, while 3HB-CoA was supplied by β-ketothiolase (PhaA) and NADPH-dependent acetoacetyl-CoA reductase (PhaB). The functional expression of these enzymes led to a production of P(LA-co-3HB) with high LA fractions (96.8 mol%). The omission of PhaA and PhaB from this pathway led to a further increase in LA fraction up to 99.3 mol%. The newly engineered C. glutamicum potentially serves as a food-grade and biomedically applicable platform for the production of poly(lactic acid)-like polyester

Elucidation of Beta-Oxidation Pathways in Ralstonia Eutropha H16 by Examination of Global Gene Expression

Ralstonia eutropha H16 is capable of growth and polyhydroxyalkanoate production on plant oils and fatty acids. However, little is known about the triacylglycerol and fatty acid degradation pathways of this bacterium. We compare whole-cell gene expression levels of R. eutropha H16 during growth and polyhydroxyalkanoate production on trioleate and fructose. Trioleate is a triacylglycerol that serves as a model for plant oils. Among the genes of note, two potential fatty acid β-oxidation operons and two putative lipase genes were shown to be upregulated in trioleate cultures. The genes of the glyoxylate bypass also exhibit increased expression during growth on trioleate. We observed that single β-oxidation operon deletion mutants of R. eutropha could grow using palm oil or crude palm kernel oil as the sole carbon source, regardless of which operon was present in the genome, but a double mutant was unable to grow under these conditions. A lipase deletion mutant did not exhibit a growth defect in emulsified oil cultures but did exhibit a phenotype in cultures containing nonemulsified oil. Mutants of the glyoxylate shunt gene for isocitrate lyase were able to grow in the presence of oils, while a malate synthase (aceB) deletion mutant grew more slowly than wild type. Gene expression under polyhydroxyalkanoate storage conditions was also examined. Many findings of this analysis confirm results from previous studies by our group and others. This work represents the first examination of global gene expression involving triacylglycerol and fatty acid catabolism genes in R. eutropha.Malaysia-MIT Biotechnology Partnership Programm

Application of a non-halogenated solvent, methyl ethyl ketone (MEK) for recovery of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(HB-co-HV)] from bacterial cells

Conventional solvent-based methods are still the most practical approaches for recovery of polyhydroxyalkanoate (PHA) polymer from cellular biomass, even though potential alternatives exist, including chemical, mechanical, and enzymatic methods. It is still necessary, however, to avoid dangerous and environmentally unfriendly solvents (e.g., chloroform and dichloromethane) in the polymer recovery process. In the work presented here, we applied various solvent systems to recover PHA from Ralstonia eutropha and recombinant Escherichia coli cells. It was demonstrated that methyl ethyl ketone (MEK) is a promising solvent for PHA recovery from bacterial cells, particularly for the copolymer poly(hydroxybutyrate-cohydroxyvalerate) [P(HB-co-HV)], exhibiting > 90% polymer recovery. Even though MEK did not solubilize PHAs to the same extent as chloroform, it can recover a comparable amount of polymer because of its processing advantages, such as the low viscosity of the MEK/PHA solution, and the lower density of MEK as compared to cellular components. MEK was found to be the best alternative, non-halogenated solvent among examined candidates for recovery of P(HB-co-HV) from cells. The MEK treatment of PHAcontaining cells further allowed us to eliminate several costly and lengthy steps in the extraction process, such as cell lysis, centrifugation, and filtration.Korea (South). Ministry of Education (Basic Science Research Program through the National Research Foundation of Korea (NRF- 2013R1A1A2A10004690))Korea Polar Research Institute (PE14030

Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(HB-co-HHx)) from butyrate using engineered Ralstonia eutropha

Polyhydroxyalkanoates (PHAs), a promising family of bio-based polymers, are considered to be alternatives to traditional petroleum-based plastics. Copolymers like poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (P(HB-co-HHx)) have been shown to exhibit favorable physical and mechanical properties, due to decreased crystallinity resulting from the presence of medium-chain-length 3-hydroxyhexanoate (3HHx) monomers. In this study, we produced P(HB-co-HHx) using engineered Ralstonia eutropha strains containing deletions of the acetoacetyl-CoA reductase (phaB) genes and replacing the native PHA synthase with phaC2 from Rhodococcus aetherivorans I24 and by using butyrate, a short-chain organic acid, as the carbon source. Although the wild-type R. eutropha did not produce P(HB-co-HHx) when grown on mixed acids or on butyrate as the sole carbon source, we are able to produce polymer containing up to 40 wt% 3HHx monomer with the aforementioned engineered R. eutropha strains using various concentrations of just butyrate as the sole carbon source. This is the first report for the production of P(HB-co-HHx) copolymer in R. eutropha using butyrate.Korea Polar Research Institute. Polar Academic Program (PAP, PD13010)Korea (South). Rural Development Administration (Project No. 010205022014

Improved Detergent-Based Recovery of Polyhydroxyalkanoates (Phas).

Extracting polyhydroxyalkanoate (PHA) polymer from bacterial cells often involves harsh conditions, including use of environmentally harmful solvents. We evaluated different detergents under various conditions to extract PHA from Ralstonia eutropha and Escherichia coli cells. Most detergents tested recovered highly pure PHA polymer from cells in amounts that depended on the percentage of polymer present in the cell. Detergents such as linear alkylbenzene sulfonic acid (LAS-99) produced a high yield of high purity polymer, and less detergent was needed compared to the amount of SDS to produce comparable yields. LAS-99 also has the advantage of being biodegradable and environmentally safe. Chemical extraction of PHA with detergents could potentially minimize or eliminate the need to use harsh organic solvents, thus making industrial PHA production a cleaner technology process.Malaysian Office of Science, Technology and Innovatio