Modeling the Interfacial Interactions between CrtS and CrtR from Xanthophyllomyces dendrorhous , a P450 System Involved in Astaxanthin Production

Xanthophyllomyces dendrorhous is a natural source of astaxanthin, a carotenoid widely used in the food industry. In this yeast, astaxanthin is synthesized from β-carotene by a cytochrome P450, CrtS, which depends on CrtR, the four-domain cytochrome P450 reductase (CPR). Although Saccharomyces cerevisiae has an endogenous CPR (ScCPR), expression of CrtS does not result in astaxanthin production unless it is coexpressed with CrtR. Assuming that CrtS could interact with the FMN-binding domain of either CrtR or ScCPR (XdFMNbd and ScFMNbd, respectively), the aim of this work was to identify possible interaction differences between these alternative complexes by protein modeling and short molecular dynamics simulations. Considering the recently proposed membrane orientation of a mammalian P450, our CrtS-CrtR model predicts that both N-terminal ends stand adjacent to the membrane plane, allowing their anchoring. Compared with the possible interface between CrtS and both FMNbd, the Xanthophyllomyces system appears to be stabilized by more saline bridges.     

Linalool from Lippia alba: study of the reproducibility of the essential oil profile and the enantiomeric purity

A new chemotype of the aromatic Verbenaceae species Lippia alba Mill. N. E. Br. from southeastern Brazil has recently been shown to have a high content of linalool in the leaf essential oil. Vegetative propagation of this chemotype was conducted at six different locations in Brazil, and the variation of the content and the optical purity of linalool in the oils were verified. Yields (0.6-0.9%, hydrodistillation), chemical composition, linalool content, and optical purity of the oils from all the plants were compared, using GC-FID, GC-MS, chiral chromatography, and retention index calculation. No plant exceeded the matrix in linalool content (46.5 to 90.7%), and the chemical profile of the oils was the same for all the samples. Purification of linalool to a content close to 100% was effected by vacuum distillation of the crude oil. Chiral analysis showed exclusively the presence of S-linalool in all the crude oils and in the distilled samples.     

N-acyl-homoserine lactones from Enterobacter sakazakii (Cronobacter spp.) and their degradation by Bacillus cereus enzymes

A chemical study of acyl-homoserine lactones (acyl-HSLs) produced by Enterobacter sakazakii resulted in the identification of three molecules: (S)-N-heptanoyl-HSL, (S)-N-dodecanoyl-HSL and (S)-N-tetradecanoyl-HSL. Mixed cultures of E. sakazakii and Bacillus cereus depleted E. sakazakii acyl-HSLs, suggesting acyl-HSL degradation by B. cereus hydrolases (hydrolysis of the lactone or amide moiety). The expression of B. cereus acyl-HSL lactonase and acyl-homoserine acylase was confirmed by monitoring the biotransformation of (S)-N-dodecanoyl-HSL into (S)-N-dodecanoyl-homoserine, dodecanoic acid and homoserine in the presence of B. cereus whole cells, using electrospray-mass spectrometry (ESI-MS).     

Effect of a Bowman-Birk proteinase inhibitor from Phaseolus coccineus on Hypothenemus hampei gut proteinases in vitro

The coffee berry borer, Hypothenemus hampei (Ferrari), is an important devastating coffee pest worldwide. Both trypsin and chymotrypsin enzyme activities from H. hampei larval midgut can be inactivated by proteinaceous enzyme-inhibitors. A serine proteinase inhibitor belonging to the Bowman-Birk class was purified from a wild accession of Phaseolus coccineus L. seeds. The inhibitor (PcBBI1) is a cysteine-rich protein that is heat-stable at alkaline pH. MALDI-TOF/MS analysis showed that PcBBI1 occurs in seeds as a monomer (8689 Da) or dimer (17,378 Da). Using in vitro inhibition assays, it was found that PcBBI1 has a high inhibitory activity against H. hampei trypsin-like enzymes, bovine pancreatic chymotrypsin, and trypsin. According to this, PcBBI1 could be a promising tool to make genetically modified coffee with resistance to coffee berry borer.     

Liming as an ameliorator of copper toxicity in black oat (Avena strigosa Schreb.)

This study aimed to assess the effect of liming as an ameliorator of copper (Cu) toxicity in black oats (Avena strigosa Schreb.) grown in soil with a sandy texture. The treatments were soil supplementation with increasing doses of lime (0.0, 0.5 and 1.0 g kg^?1) and two of Cu (0 and 50 mg kg^?1). Pre-germinated black oat seeds were sown for 30 days in the soil, subsequently, the plant height, root and shoot dry mass and concentration of total chlorophyll, chlorophylls a and b, carotenoids and total soluble carbohydrates and starch were analyzed. The dose of 50 mg kg^?1 soil Cu caused a toxic effect on black oat plant growth and development, affected the internal distribution of nutrients, decreased the leaf chlorophyll concentration and increased the leaf concentration of total soluble carbohydrates and starch. Increasing lime doses ameliorated the toxic effects of Cu.     

Generation of a Mixolab Profile After the Evaluation of the Functionality of Different Commercial Wheat Flours for Hot‐Press Tortilla Production

Refined wheat flours commercially produced by five different U.S. and Mexican wheat blends intended for tortilla production were tested for quality and then processed into tortillas through the hot‐press forming procedure. Tortilla‐making qualities of the flour samples were evaluated during dough handling, hot pressing, baking, and the first five days on the shelf at room temperature. The predominant variables that affected the flour tortilla performance were wet gluten content, alveograph W (220–303) and P/L (0.70–0.94) parameters, farinograph water absorption (57%) and stability (10.8–18.7 min), starch damage (5.43–6.71%), and size distribution curves (uniform particle distribution). Flours produced from a blend of Dark Northern Spring (80%) and Mexican Rayon (20%) wheat had the highest water absorption, and tortillas obtained from this blend showed the highest diameter and lowest thickness. The whitest and best textured tortillas were obtained from the flour milled from three hard types of Mexican wheat blend. A Mixolab profile was generated from the best tortilla flours, those produced by mills 3 and 4. The Mixolab profile showed that a good flour for hot‐press tortillas had a relatively lower absorption and short dough mix time compared with a bread flour and should have a significantly higher gluten compared with an all‐purpose flour. Compared with bread flour, the tortilla flour had higher retrogradation and viscosity values. The Mixolab profile proved to be a good preliminary test to evaluate flours for hot‐press tortillas.     

Carcass characteristics and meat quality of sheep fed alfalfa hay to replace Bermuda grass hay

Tropical Animal Health and Production - The objective of this study was to evaluate carcass characteristics, meat quality, and organ and viscera weight of forty male sheep, not castrated, initially...