Effects of heavy-ion beam irradiation on avermectin B1a and its analogues production by Streptomyces avermitilis.

The biggest challenge in anabolism research is to improve the stability and safety of microbial metabolite production on an industrial scale. One class of metabolites, avermectins, are produced by Streptomyces avermitilis. In this study, an avermectin B1a-high-producing mutant was produced using heavy ion mutagenesis and selected based on LTQ-MS and HPLC-UV method. The mutants ZJAV-Y-147 and ZJAV-Y-HS, obtained after subjecting the spores of S. avermitilis to 70 Gy of 12C6+ heavy ion irradiation, were found to best improve the avermectin B1a production (4822.23 µg/mL and 4632.17 µg/mL, respectively). These two mutants' yielded of avermectin B1a were 2-fold high than the original strains. The DNA of the original and mutant strains were analyzed by RAPD technique with four random primers after irradiated with ion beam irradiation. The results show that different high-titer S. avermitilis strains contain different genetic modifications. In addition, the mutation position, mutation type and sequence context of all mutations of aveC, aveD, aveI, aveR gene in two mutants S.avermitilis were researched, and the production of avermectin B1a and its analogues of wild-type and mutants were analyzed by fermenting 240 h, which was suggested that the partial base deletion of aveI gene may be the key sites for increasing avermectin B1a production after the 12C6+-ion irradiation. All these modifications promote increased avermectin biosynthesis, leading to multiple high-titer S. avermitilis strains. The results demonstrate that this is an effective approach to engineer S. avermitilis as a host for the biological production of commercial analogs.

In vivo blood glucose quantification using Raman spectroscopy.

We here propose a novel Raman spectroscopy method that permits the noninvasive measurement of blood glucose concentration. To reduce the effects of the strong background signals produced by surrounding tissue and to obtain the fingerprint Raman lines formed by blood analytes, a laser was focused on the blood in vessels in the skin. The Raman spectra were collected transcutaneously. Characteristic peaks of glucose (1125 cm(-1)) and hemoglobin (1549 cm(-1)) were observed. Hemoglobin concentration served as an internal standard, and the ratio of the peaks that appeared at 1125 cm(-1) and 1549 cm(-1) peaks was used to calculate the concentration of blood glucose. We studied three mouse subjects whose blood glucose levels became elevated over a period of 2 hours using a glucose test assay. During the test, 25 Raman spectra were collected transcutaneously and glucose reference values were provided by a blood glucose meter. Results clearly showed the relationship between Raman intensity and concentration. The release curves were approximately linear with a correlation coefficient of 0.91. This noninvasive methodology may be useful for the study of blood glucose in vivo.