Synthesis of poly vinyl chloride/chlorinated polypropylene-active natural substance derivatives for potential packaging materials application. Tannic acid, menthol and lipoic acid.

A novel one step route for the synthesis of tannic acid, lipoic acid and menthol functionalized polyvinyl chloride, PVC, (PVC-Tann, PVC-Lip, PVC-Mnt) and chlorinated polypropylene, PP-Cl, (PP-Mnt, PP-Lip) was applied imparting antioxidative properties to the newly-formed materials. The resulting modified polymers were characterized by stress-strain mechanical measurement, 1H NMR, gel permeation chromatography (GPC) and thermogravimetric (TGA) analysis. Linseed oil, owing to its high linolenic acid content, was used to track the autoxidation process. The chloride functional groups were reacted with the hydroxyl/carboxylic acid of the natural compounds in the presence of a base. Linseed oil was poured into the natural compound functionalized PVC covered Petri dish to undergo autoxidation under white light. Each of the PVC- and PP-Cl-based polymers was active in delaying autoxidation. The PP-Mnt series and PP-Lip polymers all delayed autoxidation by 8 days over the PP-Cl precursor material. The autoxidation process was further confirmed by monitoring peroxide formation in the exposed linseed oil samples through differential scanning calorimeter (DSC) analysis.

Sustainable bio-based antimicrobials derived from fatty acids: Synthesis, safety, and efficacy.

Some conventional sanitizers and antibiotics used in food industry may be of concerns due to generation of toxic byproducts, impact on the environment, and the emergence of antibiotic resistance bacteria. Bio-based antimicrobials can be an alternative to conventional sanitizers since they are produced from renewable resources, and the bacterial resistance to these compounds is of less concern than those of currently used antibiotics. Among the bio-based antimicrobial compounds, those produced via either fermentation or chemical synthesis by covalently or electrovalently attaching specific moieties to the fatty acid have drawn attention in recent years. Disaccharide, arginine, vitamin B1, and phenolics are linked to fatty acids resulting in the production of sophorolipid, lauric arginate ethyl ester, thiamin dilauryl sulfate, and phenolic branched-chain fatty acid, respectively, all of which are reported to exhibit antimicrobial activity by targeting the cell membrane of the bacteria. Also, studies that applied these compounds as food preservatives by combining them with other compounds or treatments have been reviewed regarding extending the shelf life and inactivating foodborne pathogens of foods and food products. In addition, the phenolic branched-chain fatty acids, which are relatively new compounds compared to the others, are highlighted in this review.

Synthesis of a novel tannic acid-functionalized polypropylene as antioxidant active-packaging materials.

This work focuses on the synthesis of novel tannin-functionalized polypropylene copolymers that are designed to inhibit the oxidation of vegetable oils for potential use as packaging materials. An empty glass Petri dish (control), a chlorinated polypropylene-coated glass Petri dish (control) and a series of the tannin-functionalized polypropylene coated glass Petri dishes overlaid with linseed oil were exposed to air and additional white light. Oligomerization of the oxidized linseed oil was assessed by measuring the flow properties of the exposed oil using a viscometer. The antioxidant effect of the tannic acid grafted polypropylene copolymers (PP-Tann) retarded oligomerization of the linseed oil. The molar mass of the linoleic acid overlaid onto the PP-Tann films was the lowest among the tested samples after each time period indicating that tannin-grafted polypropylene may be a promising packaging material for vegetable oils.

Burkholderia sacchari DSM 17165: A source of compositionally-tunable block-copolymeric short-chain poly(hydroxyalkanoates) from xylose and levulinic acid.

Burkholderia sacchari was used to produce poly-3-hydroxybutyrate-co-3-hydroxyvalerate block copolymers from xylose and levulinic acid. Levulinic acid was the preferred substrate resulting in 3-hydroxyvalerate (3HV) contents as high as 95 mol% at 24 h. The 3HB:3HV ratios were controlled by the initial levulinic acid media concentration and fermentation length. Higher levulinic acid concentrations and longer durations, resulted in polymers with two glass transition temperatures, each approximating those associated with poly-3HB and poly-3HV. 13C NMR confirmed the presence of high concentrations of 3HB-3HB and 3HV-3HV homopolymeric dyads, while mass spectrometry of the partial hydrolysis products did not conform to Bernoullian statistics for randomness, confirming block sequences. MS/MS analysis of specific oligomers showed the mass-loss of 86 amu (a 3HB unit) and 100 amu (a 3HV unit) attesting to some randomness within the polymers. This study verifies the potential for producing Poly-3HB-block-3HV copolymers from inexpensive biorenewable feedstocks without sequential addition of carbon sources.

Characterization of growth inhibition of oral bacteria by sophorolipid using a microplate-format assay.

Sophorolipid (SL) is a class of glycolipid biosurfactant produced by yeast and has potent antimicrobial activity against many microorganisms. In this paper, a microplate-based method was developed to characterize the growth inhibition by SL on five representative species of caries-causing oral bacteria. Bacterial growth on microplate in the absence and presence of varying concentrations of SL was continuously monitored by recording the absorbance at 600nm of the cultures using a microplate reader. The results showed that SL completely inhibited the growth of the Lactobacilli at ≥1mg/ml and the Streptococci at much lower concentrations of ≥50µg/ml. More importantly, we further defined the mechanism of antimicrobial activity of SL by analyzing the pattern of the cell growth curves. SL at sublethal concentrations (<1mg/ml) is bactericidal towards the Lactobacilli; it lengthens the apparent cell-doubling time (Td) and decreases the final cell density (as indicated by A600nm) in a concentration-dependent manner. Against the oral Streptococci, on the other hand, SL at sublethal concentrations (<50µg/ml) is bacteriostatic; it delays the onset of cell growth in a concentration-dependent fashion, but once the cell growth is commenced there is no noticeable adverse effect on Td and the final A600nm. Scanning electron microscopic (SEM) study of L. acidophilus grown in sublethal concentration of SL reveals extensive structural damage to the cells. S. mutans grown in sublethal level of SL did not show morphological damage to the cells, but numerous protruding structures could be seen on the cell surface. At the respective lethal levels of SL, L. acidophilus cells were lysed (at 1mg/ml SL) and the cell surface structure of S. mutans (at 130µg/ml SL) was extensively deformed. In summary, this paper presents the first report on a detailed analysis of the effects of SL on Lactobacilli and Streptococci important to oral health and hygiene.

Preparation of starch-poly-glutamic acid graft copolymers by microwave irradiation and the characterization of their properties.

Graft copolymers of waxy maize starch and poly-γ-glutamic acid (PGA) were produced in an aqueous solution using microwave irradiation. The microwave reaction conditions were optimized with regard to temperature and pH. The temperature of 180°C and pH7.0 were the best reaction conditions resulting in a PGA graft of 0.45% based on nitrogen analysis. The average graft content and graft efficiency for the starch-PGA graft copolymer prepared at 180°C and pH7.0 were 4.20% and 2.73%, respectively. The starch-PGA graft copolymer produced at 180°C and pH7.0 could absorb more than 20 times its own weight amount of water and form a gel. The preliminary rheology study revealed that the starch-PGA graft copolymer gel exhibited viscoelastic solid behavior while the control sample of waxy starch showed viscoelastic liquid behavior.

High-titer production and strong antimicrobial activity of sophorolipids from Rhodotorula bogoriensis.

Rhodotorula bogoriensis produces sophorolipids (SLs) that contain 13-hydroxydocosanoic acid (OH-C22 ) as the lipid moiety. A systematic study was conducted to further understand the fermentative production of SLs containing OH-C22 (C22 -SL) by R. bogoriensis. Shake-flask studies showed that R. bogoriensis consumed glucose at a slow pace. HPLC analysis of the C22 -SL products from shake-flask fermentations at different glucose concentrations showed a correlation between glucose depletion and the extent of C22 -SL deacetylation. A large-scale bioreactor fermentation resulted in the isolation of C22 -SL at a volumetric product yield of 51 g/L. HPLC analysis of C22 -SL product from the bioreactor fermentation corroborated the finding that glucose depletion correlated with extensive deacetylation of C22 -SL. The antimicrobial activity of C22 -SL was established for the first time to be stronger than the C18 -SL from Candida bombicola against Propionibacterium acnes in a plate assay.

Dirhamnose-lipid production by recombinant nonpathogenic bacterium Pseudomonas chlororaphis.

We previously discovered that Pseudomonas chlororaphis NRRL B-30761 produces monorhamnolipids (R1Ls) with predominantly 3-hydroxydodecenoyl-3-hydroxydecanoate (C12:1-C10) or 3-hydroxydodecanoyl-3-hydroxydecanoate (C12-C10) as the lipid moiety under static growth conditions only. We have now cloned, sequenced, and analyzed in silico the gene locus of NRRL B-30761 containing the putative coding sequences of rhamnosyltransferase chain A (rhlA Pch , 894 bps), rhamnosyltransferase chain B (rhlB Pch , 1272 bps), and N-acyl-homoserine lactone-dependent transcriptional regulatory protein (rhlR Pch , 726 bps). The putative gene products RhlAPch (297 amino acid residues or a.a.), RhlBPch (423 a.a.), and RhlRPch (241 a.a.) only have between 60 and 65% a.a. identities to their respective closest matched homologs in P. aeruginosa. Polymerase chain reaction (PCR)-based assay did not detect the presence of rhamnosyltransferase C gene (rhlC) in P. chlororaphis, suggesting a genetic basis for the lack of dirhamnose-lipid (R2L) synthesis in this organism. We thus genetically constructed an R2L-synthesizing P. chlororaphis by expressing a rhamnosyltransferase C (rhlC) gene of P. aeruginosa using an expression vector (pBS29-P2-gfp) containing a Pseudomonas syringae promoter. The R2L/R1L ratio is 2.4 in the rhamnolipid (RL) sample isolated from the genetically engineered (GE) P. chlororaphis [pBS29-P2-rhlC], in contrast to undetectable R2L in the GE P. chlororaphis [pBS29-P2-gfp] control cells based on LC-MS analysis. The critical micelle concentrations of the R2L and R1L samples from GE P. chlororaphis [pBS29-P2-rhlC] and the control [pBS29-P2-gfp] cells were ca. 0.1 mM, and their minimum surface tensions were ca. 26 mN/m with no significant difference.

Methanol-induced chain termination in poly(3-hydroxybutyrate) biopolymers: molecular weight control.

A systematic study was performed to demonstrate the impact of methanol (MeOH) on poly(3-hydroxybutyrate) (PHB) synthesis and molecular weight (MW). Glycerine was used as the primary carbon source with varying concentrations of MeOH. Methanol retarded but did not completely inhibit growth and PHB production in Pseudomonas oleovorans. Proton NMR analysis revealed that the PHB polymers were end-capped with methoxy chemical groups causing MW reductions. The MW decreases were contingent upon the initial MeOH media concentration and the duration of the fermentations. The largest impact occurred at an initial MeOH concentration of 0.10% (w/v) where the number average molecular weights (Mn) decreased by 39%, 55%, and 72% in the 48, 72 and 96 h cultures, respectively. Diffusion ordered NMR spectroscopy revealed a diffusivity (D) increase in the smaller molecular weight polymers with the PHB synthesized in the presence of 0.85% MeOH (72 h post-inoculation) having a D value of 0.66×10(-10) m2/s. Diffusivity increases indicate a reduction in hydrodynamic radii (Rhz) consistent with shorter chain-lengths. Crude glycerine from the biodiesel production process has been used as an inexpensive fermentation feedstock for polyhydroxyalkanoate (PHA) synthesis but its composition is facility-dependent. This information will be vital to tailor PHA properties to specific applications.

Glycerine and levulinic acid: renewable co-substrates for the fermentative synthesis of short-chain poly(hydroxyalkanoate) biopolymers.

Glycerine (a biodiesel co-product) and levulinic acid (a pulp and paper co-product) were used as co-substrates for the fermentative synthesis of short-chain polyhydroxyalkanoate (sc-PHA) biopolymers with tunable monomer and molecular weight characteristics. Pseudomonas oleovorans NRRL B-14682 utilized glycerine alone to produce poly(3-hydroxybutyrate) (PHB). When levulinic acid was added to the media at shake-flask scale in concentrations ≤0.6 wt.%, poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHB/V) copolymers were produced with 3-HV contents ranging from 37 to 97 mol%; a glycerine:levulinic acid ratio of 0.2%:0.8% (w/v) resulted in poly(3-hydroxyvalerate) (PHV). Ten-liter batch fermentations using glycerine:levulinic acid ratios of 1%:0, 0.75%:0.25%, 0.5%:0.5% and 0.25%:0.75% (w/v) resulted in PHB, P(73%-3HB-co-27%-3HV), P(30%-3HB-co-70%-3HV) and PHV with increasing number average molecular weights (×10(3) g/mol) of 328, 511, 728 and 1330, respectively, owing to glycerine-based chain termination. These results provide a novel means by which glycerine and levulinic acid can be used collectively to produce an array of distinct sc-PHA biopolymers.

Cloning, sequencing, and characterization of lipase genes from a polyhydroxyalkanoate (PHA)-synthesizing Pseudomonas resinovorans.

Lipase (lip) and lipase-specific foldase (lif) genes of a biodegradable polyhydroxyalkanoate (PHA)-synthesizing Pseudomonas resinovorans NRRL B-2649 were cloned using primers based on consensus sequences, followed by polymerase chain reaction-based genome walking. Sequence analyses showed a putative Lip gene product (314 amino acids, a.a.) with its catalytic active site (Ser(111), Asp(258), and His(280)) identified. The foldase lif gene that is located 55 bp downstream of lip codes for a putative Lif (345 a.a.). To verify the biological function of the cloned lip gene for lipase expression in P. resinovorans, we constructed a lip knock-out mutant (lip::Tn5) by transposon insertion. Complementation of the lip knock-out P. resinovorans mutant with a lipase expression plasmid (pBS29-P2-lip) was performed, and its effect on lipase expression was investigated. The wild-type P. resinovorans and the lip::Tn5[pBS29-P2-lip] recombinant (but not the lip::Tn5 mutant) showed fluorescence on rhodamine B plates indicative of lipase activity. The wild type exhibited extracellular lipase activity when grown on medium containing triacylglycerol substrates (tallow, olive oil, and tributyrin) as sole carbon sources, but the lip::Tn5 mutant did not show such activity. Lipase activity of various strains was also confirmed by TLC analysis of the composition of acylglycerols and free fatty acid in the extracts of the spent culture medium. We further found that tributyrin was more effective than olive oil in inducing lipase expression in P. resinovorans.

Rendered-protein hydrolysates for microbial synthesis of cyanophycin biopolymer.

Cyanophycin is a poly(arginyl-aspartate) biopolymer produced and stored intracellularly by bacteria. Cyanophycin has been proposed as a renewable replacement for petrochemical-based industrial products. An abundant source of amino acids and nitrogen such as in the form of protein hydrolysates is needed for the biosynthesis of cyanophycin. Rendered proteins are largely used as a feed supplement in animal husbandry and aquaculture. New uses would expand the market size of this class of protein coproducts. We prepared and thoroughly characterized the hydrolysates of meat and bone meal, and proceeded to demonstrate for the first time that these hydrolysates could be used in the fermentative production of cyanophycin. Using the enzyme-hydrolyzed meat and bone meal preparation, we obtained crude cyanophycin product at 33-35% level of that produced using the reference casamino acids in both shake-flask and 10-L bioreactor fermentation studies. Polyacrylamide-gel electrophoresis of the cyanophycin under denaturing conditions showed the molecular weight of the isolated polyamide at 24kDa. Our results open a new avenue for the utilization of rendered protein coproducts to produce the cyanophycin biopolymer.

Biopolymer scaffolds for use in delivering antimicrobial sophorolipids to the acne-causing bacterium Propionibacterium acnes.

Sophorolipids (SLs) are known to possess antimicrobial properties towards many species (particularly Gram-positive, or Gram(+)) of bacteria. However, these properties can only be exerted if the SLs can be introduced to the bacterial cells in an acceptable manner. Propionibacterium acnes is the common bacterial cause of acne. It is a Gram(+) facultative anaerobe that is susceptible to the antimicrobial effects of SLs. In this study we demonstrated that different biopolymer matrices could be used to produce SL composite films that exert various antimicrobial efficiencies against P. acnes. Increasing SL concentrations in poly-3-hydroxybutyrate (PHB) and PHB-co-10%-3-hydroxyhexanoate (PHB/HHx) resulted in noticeably improved (PHB/HHx was best) antimicrobial activity based on the size of the zones of inhibition using an overlay plating technique on synthetic growth medium. However, increasing concentrations of SLs in PHB and PHB/HHx films also increased film opacity, which diminishes the appeal for use especially in visible (facial) areas. Pectin and alginate improved the transparent character of SL composite films while also acting as successful carriers of SLs to P. acnes. The lactone form of the SLs proved to exhibit the best antimicrobial action and in concert with either pectin or alginate biopolymers provided a comparatively transparent, successful means of utilizing SLs as a renewable, environmentally benign anti-acne solution.

A new shuttle vector for gene expression in biopolymer-producing Ralstonia eutropha.

Ralstonia eutropha (formerly Alcaligenes eutrophus) is a fascinating microorganism with a great scientific importance and an immense commercial potential. A new genetic transformation system for the organism would greatly facilitate the biological study and molecular engineering of this organism. We report here a versatile gene expression method for the genetic engineering of R. eutropha. This method, based on a simplified electroporation protocol, uses a recombinant plasmid, pBS29-P2, containing a Pseudomonas syringae promoter (P2) and two antibiotic-resistance markers (i.e., genes coding for kanamycin (Km)- and tetracycline (Tc)-resistance). Using this method, we successfully achieved transformation of wild-type R. eutropha and its poly(hydroxyalkanoate)-negative mutant, R. eutropha PHB(-)4, with various pBS29-P2-based recombinants. A transformation frequency as high as 4x10(3) Km-resistance colonies/mug DNA was obtained per electroporation experiment. We further demonstrated the successful expression of a heterologous gene coding for green-fluorescent-protein by fluorescence measurement. In addition, our results indicated the expression of a truncated but active Streptomyces coelicolor alpha-galactosidase in R. eutropha.

The influence of increasing media methanol concentration on sophorolipid biosynthesis from glycerol-based feedstocks.

Candida bombicola, a known producer of sophorolipids (SLs; glycolipid surfactants), was grown on glycerol and oleic acid with up to 1.5% (v/v) methanol in the fermentation growth media to assess the effects of methanol presence on SL synthesis and structural distribution. Increasing methanol concentrations had little effect on the growth of the organism resulting in average cell dry weights (CDW; after SL separation) of 20.8 ± 0.7 g/l between 0 and 1.5% methanol. However, increasing methanol concentrations decreased SL production by 56% (from 12.7 to 5.6 g/l at 1.5% methanol) which translated to SL yields on a cellular basis of between 0.60 g SL/g cells (in the absence of methanol) to 0.27 g SL/g cells (in the presence of 1.5% methanol). LC/MS revealed that increased methanol concentrations also resulted in larger concentrations (up to 20 mol%) of free acid SLs but had little effect on the ratios of diacetylated SL lactones synthesized with palmitic acid (4 mol%), linoleic acid (3 mol%), oleic acid (80 mol%), and stearic acid (13 mol%) as the hydrophobic moieties.

Methyl-branched poly(hydroxyalkanoate) biosynthesis from 13-methyltetradecanoic acid and mixed isostearic acid isomer substrates.

Pseudomonas resinovorans, a known medium-chain-length (mcl-) poly(hydroxyalkanoate) (PHA) producer, was grown on 13-methyltetradecanoic acid (13-MTDA) and a mixture of isostearic acid (IA) isomers to produce methyl-branched mcl-PHA polymers. Shake-flask experiments revealed polymer productivities (the percent of the cell mass that is polymer) of 31 +/- 1% (n = 3) and 23 +/- 3% (n = 3) when grown in 13-MTDA and IA, respectively. Monomer content was determined by a combination of gas chromatography/mass spectrometry (GC/MS) of the acid hydrolyzed, silylated methyl esters, and nuclear magnetic resonance spectroscopy. Results showed that the mcl-PHA polymer derived from 13-MTDA was primarily composed of 3-hydroxy-7-methyloctanoic acid and 3-hydroxy-9-methyldecanoic acid (67 and 16 mol% by GC/MS, respectively). In contrast, the mcl-polymers synthesized from the IA isomeric mixture were more complex, containing both even and odd chain-length monomers as well as varying distributions of methyl-branched derivatives. The PHA distributions among the C8, C10, C12, and C14 carbon chain-length monomers included three isomers of C8, five isomers of C10, seven isomers of C12, and nine isomers of C14 each containing one linear-chain derivative and n-6 methyl-branched derivatives where n equals the total number of carbon atoms in each monomer unit (C8-C14).

Altered composition of Ralstonia eutropha poly(hydroxyalkanoate) through expression of PHA synthase from Allochromatium vinosum ATCC 35206.

The class III poly(hydroxyalkanoate) synthase (PHAS) genes (phaC and phaE) of a photosynthetic bacterium, Allochromatium vinosum ATCC 35206, were cloned, sequenced and expressed in a heterologous host. PCR coupled with a chromosomal gene-walking method was used to clone and subsequently sequence the contiguous phaC (1,068 bps) and phaE (1,065 bps) genes of A. vinosum ATCC 35206. BLASTP search of protein databases showed that the gene-products of phaC and phaE are different (<66% identities) from the previously reported class III PHASs such as those of A. vinosum DSM180. Domain analysis revealed the presence of a conserved alpha/beta-hydrolase fold in PhaC, the putative gene-product of phaC. Upon electroporation of a poly(hydroxybutanoate) (PHB)-negative mutant of Ralstonia eutropha PHB(-)4 with a shuttle plasmid pBHR1 containing the newly cloned phaC and phaE genes, the bacteria resumed the synthesis of PHB, albeit at a low level (4-5% of the cell dry wt) due to kanamycin selection pressure. We further showed that the recombinant strain grown in kanamycin-containing culture medium synthesized a blend of PHA that also contains a high content of 3-hydroxyoctanoate and 3-hydroxydecanoate as its repeat-unit monomers. Genomic analysis suggested the existence of two PHA synthase genes in R. eutropha. The results of this study not only make available a phylogenetically diverse type III phaC and phaE genes, but also confirm through kanamycin selection pressure the existence of multiple PHA biosynthesis systems in R. eutropha.

Property control of sophorolipids: influence of fatty acid substrate and blending.

Sophorolipids (SLs) were synthesized by fed-batch fermentation of Candida bombicola on glucose and either palmitic acid (SL-p), stearic acid (SL-s), oleic acid (SL-o) or linoleic acid (SL-l) and the structural distribution accurately determined by atmospheric pressure chemical ionization-mass spectrometry (APCI-MS). The surfactant properties, including critical micelle concentration (CMC), minimum surface tension (min.ST) and oil-water interfacial tension (IFT) were measured by tensiometry. Minimum STs of 35-36 mN/m were obtained regardless of the substrate while IFTs ranged from 3-5 mN/m with the exception of SL-l, which had an IFT of 7 mN/m. The largest disparity occurred in the CMC values, which ranged from 35 ppm for SL-s to 250 ppm for SL-l. By manually mixing these four SLs in different ratios, it was possible to better control the CMC values without affecting the min.ST or IFT, which will prove beneficial as new applications for SLs are established.

Genetic organization of pha gene locus affects phaC expression, poly(hydroxyalkanoate) composition and granule morphology in Pseudomonas corrugata.

The complete sequence of the pha locus responsible for the biosynthesis of poly(hydroxyalkanoates) (PHAs) in Pseudomonas corrugata 388 was determined. As with the other known pseudomonad pha gene loci, the one in P. corrugata 388 also consists of phaC1 (1,680 bps; PHA synthase 1), phaZ (858 bp; PHA depolymerase) and phaC2 (1,683 bp; PHA synthase 2) genes. A BLAST search showed that the nucleotide sequences of these genes and the amino-acid sequences of their respective gene products are homologous to those of P. corrugata CFBP5454 and P. mediterranea CFBP5447. A putative intrinsic transcription terminator consisting of a dyad symmetry (24 bp; Delta G = -41.8 kcals) that precedes a stretch of dA residues was located in the phaC1-phaZ intergenic region. P. corrugata mutant-clones XI 32-1 and XI 32-4 were constructed in which this intergenic region was replaced with a selectable kanamycin-resistance gene. These mutant clones when grown on oleic acid for 48 h showed 4.7-to 7.0-fold increases of phaC1 and phaC2 relative expression in comparison to the initial inoculants, whereas the parental strain showed only 1.2- to 1.4-fold increases. Furthermore, in comparison to parental P. corrugata with only a few large PHA inclusion bodies, the mutants grown on oleic acid produce numerous smaller PHA granules that line the periphery of the cells. With glucose as a substrate, XI 32-1 and XI 32-4 clones produce mcl-PHA with a high content (26-31 mol%) of the mono-unsaturated 3-hydroxydodecenoate as a repeat-unit monomer. Our results show for the first time the effects of the phaC1-phaZ intergenic region on the substrate-dependent temporal expression of phaC1 and phaC2 genes, the repeat-unit composition of mcl-PHA, and the morphology of the PHA granules.

Simplified soy molasses-based medium for reduced-cost production of sophorolipids by Candida bombicola.

A simplified medium containing only soy molasses and oleic acid as ingredients was developed for the production of sophorolipids (SLs) from Candida bombicola. We achieved a product yield of 53 +/- 3 g of purified sophorolipids per liter of starting culture volume, which is 71 +/- 4% of the yield obtained with growth medium that also additionally contains the costly yeast extract and urea as nitrogen source. The large majority of the SL components existed in the lactone form (87%), and the predominant component is SL containing (omega-1)-hydroxyoleic acid as the lipid moiety. The study demonstrated for the first time the usefulness of the low-value soy molasses as a combined nitrogen- and carbon-source for SL production at a reduced cost.