Herein, we reveal a second quorum-sensing system produced by Cronobacter sakazakii. A cyclo(l-Pro–l-Leu) diketopiperazine, detected in pure and mixed cultures of C. sakazakii and Bacillus cereus explains the coexistence of both in the same industrial environments. The molecule was identified by gas chromatography–mass spectrometry (GC–MS), 1H, and 13C NMR, including 2D NMR (correlation spectroscopy, heteronuclear multiple bond correlation, and heteronuclear single quantum correlation), and the absolute configuration was compared with that of four synthetic standards produced by solid phase peptide synthesis using a chiral column on a GC–flame ionization detection. This article provides a new method to determine the absolute configuration of cyclo(Pro–Leu) diketopiperazine replacing the joint use of 1H NMR and Marfey’s method.

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The effects of soybean and castorbean meals were evaluated separately, and in combinations at different ratios, as substrates for lipase production by Botryosphaeria ribis EC-01 in submerged fermentation using only distilled water. The addition of glycerol analytical grade (AG) and glycerol crude (CG) to soybean and castorbean meals separately and in combination, were also examined for lipase production. Glycerol-AG increased enzyme production, whereas glycerol-CG decreased it. A 2(4) factorial design was developed to determine the best concentrations of soybean meal, castorbean meal, glycerol-AG, and KH2PO4 to optimize lipase production by B. ribis EC-01. Soybean meal and glycerol-AG had a significant effect on lipase production, whereas castorbean meal did not. A second treatment (2(2) factorial design central composite) was developed, and optimal lipase production (4,820 U/g of dry solids content (ds)) was obtained when B. ribis EC-01 was grown on 0.5 % (w/v) soybean meal and 5.2 % (v/v) glycerol in distilled water, which was in agreement with the predicted value (4,892 U/g ds) calculated by the model. The unitary cost of lipase production determined under the optimized conditions developed ranged from US$0.42 to 0.44 based on nutrient costs. The fungal lipase was immobilized onto Celite and showed high thermal stability and was used for transesterification of soybean oil in methanol (1:3) resulting in 36 % of fatty acyl alkyl ester content. The apparent K m and V max were determined and were 1.86 mM and 14.29 μmol min(-1) mg(-1), respectively.

Said de Sousa (IC)*, José E. do N. Fontes (PG), Matheus R. Bofinger (PG), Anita J. Marsaioli (PQ) Resumo As 2,5-dicetopiperazinas são moléculas utilizadas para comunicação bacteriana, ou quorum-sensing (QS). Enterobacter sakazakii e Bacillus cereus foram cultivadas isoladamente e em conjunto para avaliação da comunicação que ambas exercem e observou-se que ambas produziam a molécula Ciclo(Pro-Leu) para comunicação. A molécula foi então identificada e caracterizada utilizando-se de técnicas espectrométricas (CG-MS) e espectroscópicas (RMN de ¹H, ¹³C e 2D).