Dorselyn C. Chapital

Metabolic engineering of Saccharomyces cerevisiae for production of novel lipid compounds

Abstract. The yeast Saccharomyces cerevisiae has been modified successfully for production of numerous metabolites and therapeutic proteins through metabolic engineering, but has not been utilized to date for the production of lipid-derived compounds. We developed a lipid metabolic engineering strategy in S. cerevisiae based upon culturing techniques that are typically employed for studies of peroxisomal biogenesis; cells were grown in media containing fatty acids as a sole carbon source, which promotes peroxisomal proliferation and induction of enzymes associated with fatty acid β-oxidation. Our results indicate that growth of yeast on fatty acids such as oleate results in extensive uptake of these fatty acids from the media and a subsequent increase in total cellular lipid content from 2% to 15% dry cell weight. We also show that co-expression of plant fatty acid desaturases 2 and 3 (FAD2 and FAD3), using a fatty acid-inducible peroxisomal gene promoter, coupled the processes of fatty acid uptake with the induction of a new metabolic pathway leading from oleic acid (18:1) to linolenic acid (18:3). Finally, we show that cultivation of yeast cells in the presence of triacylglycerols and exogenously supplied lipase promotes extensive incorporation of triglyceride fatty acids into yeast cells. Collectively, these results provide a framework for bioconversion of low-cost oils into value-added lipid products.

Chinese melon (Momordica charantia L.) seed: composition and potential use

Chinese melon (Momordica charantia L.), also known as bitter gourd, is a tropical crop, grown throughout Asian countries for use as food and medicinals. In 1993, four cultivars of Chinese melon were grown in Mississippi and the seeds were collected. Oil contents of the seeds ranged from 41 to 45% and the oils contained 63–68% eleostearic acid and 22–27% stearic acid. Industrially important tung oil, a “fast-drying oil” used in paints and varnishes, contains 90% eleostearic and 2–3% stearic acid. The ratio of stearic to eleostearic in Chinese melon seed oil is ten times greater than that in tung oil. The higher ratio should reduce the rate of drying and crosslinking and could be advantageous in the paint industry. The defatted meals contained 52–61% protein and would be a good source of methionine.

Identification of a subtilisin-like protease in seeds of developing tung fruits

Summary A subtilisin-like protease was identified in the low-density microsomal fraction of developing tung seeds. The abundance of the protease changed significantly throughout seed development and showed a direct temporal correlation with accumulation of storage oil. Once storage oil synthesis was complete, the abundance of the protease decreased dramatically. Determination of the N-terminal amino acid sequence revealed that the protease sequence began just after a conserved pro-domain cleavage site, suggesting that the protease was fully active in developing tissue. Extraction of microsomal membranes with high salt or pH demonstrated that the protease was not a membrane-anchored protein.

Adhesive properties of water-washed cottonseed meal on four types of wood

Abstract The interest in natural product-based wood adhesives has been steadily increasing due to the environmental and sustainable concerns of petroleum-based adhesives. In this work, we reported our research on the utilization of water-washed cottonseed meal (WCM) as wood adhesives. The adhesive strength and water resistance of WCM adhesive preparations on poplar, Douglas fir, walnut, and white oak wood veneers were tested with press temperatures of 80, 100, and 130 °C. Our data indicated that raising the hot press temperature from 80 to 100–130 °C greatly increased the bonding strength and water resistance of the WCM adhesives. The general trend of the adhesive strength of WCM on the four wood species was Douglas fir > poplar ≈ white oak > walnut. The rough surface of Douglas fir with tipping features could enhance the mechanical interlocking between the wood fibers and adhesive slurry, contributing to the high adhesive strength. The dimensional swelling of the bonded wood pairs due to water soaking was in the order: thickness > width (i.e. perpendicular to the wood grain) > length (i.e. parallel to the wood grain). The greatest dimensional changes were observed in Douglas fir specimens. However, the highest decrease in adhesive strength by water soaking was with poplar wood specimens. These observations suggested that the wood dimensional changes were not dominant factors on water weakening the bonding strength of these wood pairs.

Microwave heating of cottonseed: A pilot plant study

Two cottonseed samples (50 kg) were exposed to microwaves at 45 KW and 2450 MHz for 4 min in an air temperature of 94°C. The final temperature of the seed was 76°C, and the treatment caused a 20% reduction in seed moisture. Examination of the seed immediately after microwave heating (MWH) indicated no differences in the total or soluble protein content of the meal or the content, quality or color of the oil as compared with unheated seed. There was some reduction in the quality of the oil from unheated seed during storage, but only minimal reduction in the quality of the oil from the MWH samples. Protein quality deteriorated in all samples; however, the deterioration was reduced in the microwave-heated seed.