A bacterial biosynthetic pathway for methylated furan fatty acids.

Fatty acids play many important roles in cells and also in industrial processes. Furan fatty acids (FuFAs) are present in the lipids of some plant, fish, and microbial species and appear to function as second messengers in pathways that protect cells from membrane-damaging agents. We report here the results of chemical, genetic, and synthetic biology experiments to decipher the biosynthesis of the monomethylated FuFA, methyl 9-(3-methyl-5-pentylfuran-2-yl) nonanoate (9M5-FuFA), and its dimethyl counterpart, methyl 9-(3,4-dimethyl-5-pentylfuran-2-yl) nonanoate (9D5-FuFA), in two α-proteobacteria. Each of the steps in FuFA biosynthesis occurs on pre-existing phospholipid fatty acid chains, and we identified pathway intermediates and the gene products that catalyze 9M5-FuFA and 9D5-FuFA synthesis in Rhodobacter sphaeroides 2.4.1 and Rhodopseudomonas palustris CGA009. One previously unknown pathway intermediate was a methylated diunsaturated fatty acid, (10E,12E)-11-methyloctadeca-10,12-dienoic acid (11Me-10t,12t-18:2), produced from (11E)-methyloctadeca-11-enoic acid (11Me-12t-18:1) by a newly identified fatty acid desaturase, UfaD. We also show that molecular oxygen (O2) is the source of the oxygen atom in the furan ring of 9M5-FuFA, and our findings predict that an O2-derived oxygen atom is incorporated into 9M5-FuFA via a protein, UfaO, that uses the 11Me-10t,12t-18:2 fatty acid phospholipid chain as a substrate. We discovered that R. palustris also contains a SAM-dependent methylase, FufM, that produces 9D5-FuFA from 9M5-FuFA. These results uncover the biochemical sequence of intermediates in a bacterial pathway for 9M5-FuFA and 9D5-FuFA biosynthesis and suggest the existence of homologs of the enzymes identified here that could function in FuFA biosynthesis in other organisms.

Perceptions and performance using computer-based testing: One institution's experience.

BACKGROUND AND PURPOSE: The purpose of this study was to evaluate student and faculty perceptions of the transition to a required computer-based testing format and to identify any impact of this transition on student exam performance. EDUCATIONAL ACTIVITY AND SETTING: Separate questionnaires sent to students and faculty asked about perceptions of and problems with computer-based testing. Exam results from program-required courses for two years prior to and two years following the adoption of computer-based testing were compared to determine if this testing format impacted student performance. FINDINGS: Responses to Likert-type questions about perceived ease of use showed no difference between students with one and three semesters experience with computer-based testing. Of 223 student-reported problems, 23% related to faculty training with the testing software. Students most commonly reported improved feedback (46% of responses) and ease of exam-taking (17% of responses) as benefits to computer-based testing. Faculty-reported difficulties were most commonly related to problems with student computers during an exam (38% of responses) while the most commonly identified benefit was collecting assessment data (32% of responses). Neither faculty nor students perceived an impact on exam performance due to computer-based testing. An analysis of exam grades confirmed there was no consistent performance difference between the paper and computer-based formats. DISCUSSION AND SUMMARY: Both faculty and students rapidly adapted to using computer-based testing. There was no evidence that switching to computer-based testing had any impact on student exam performance.