Biotransformation of isoeugenol catalyzed by growing cells of Pseudomonas putida

Abstract

Growing cells of Pseudomonas putida transformed isoeugenol after 5 days of incubation to give mainly vanillin, eugenol, 4-(E)-(3-hydroxyprop-1-enyl)-2-methoxyphenol and the dimeric molecule (+)-4-[2,3-dihydro-7-methoxy-3-methyl-5-(E)-(1-propenyl)-2-benzofuranyl]-2-methoxyphenol (licarin A). The formation of the latter compound from isoeugenol by biotransformation with P. putida is reported here for the first time.     

Production of vanillin from ferulic acid using recombinant strains ofEscherichia coli

Vanillin is one of the world's principal flavoring compounds, and is used extensively in the food industry. The potential vanillin production of the bacteria was compared to select and clone genes which were appropriate for highly productive vanillin production byE. coli. Thefcs (feruloyl-CoA synthetase) andech (enoyl-CoA hydratase/aldolase) genes cloned fromAmycolatopsis sp. strain HR104 andDelftia acidovorans were introduced to pBAD24 vector with P_BAD promoter and were named pDAHEF and pDDAEF, respectively. We observed 160 mg/L vanillin production withE. coli harboring pDAHEF, whereas 10 mg/L of vanillin was observed with pDDAEF. Vanillin production was optimized withE. coli harboring pDAHEF. Induction of thefcs andech genes from pDAHEF was optimized with the addition of 13.3 mM arabinose at 18 h of culture, from which 450 mg/L of vanillin was produced. The feeding time and concentration of ferulic acid were also optimized by the supplementation of 0.2% ferulic acid at 18 h of culture, from which 500 mg/L of vanillin was obtained. Under the above optimized condition of arabinose induction and ferulic acid supplementation, vanillin production was carried out with four different types of media, M9, LB, 2YT, and TB. The highest vanillin production, 580 mg/L, was obtained with LB medium, a 3.6 fold increase in comparison to the 160 mg/L obtained before the optimization of vanillin production.     

Purification and Characterization of Carboxypeptidases from the Sarcocarp of Watermelon

Two kinds of carboxypeptidases (F–I, F–II) were purified from the sarcocarp of watermelon (Citrullus vulgaris, var. Shimao). F–I was not purified to homogeneity. F–II was homogeneous on ultracentrifugal analysis, but a trace of impurity was detected at high concentrations by disc electrophoresis.

F–I was optimally active and stable at pH 5.0~5.5 and was strongly inhibited by DFP and HgCl₂, but not by EDTA. The molecular weight and isoelectric point were 89,000 and 4.4, respectively.

F–II was optimally active at pH 5.0 ~ 5.5 and was most stable at pH 5.5 ~ 7.0. It was completely inhibited by DFP and HgCl₂, but not by EDTA and 1, 10-phenanthroline, and it hydrolyzed an oligopeptide containing proline, glutamic acid, lysine and several neutral amino acids, sequentially from the C-terminal. The molecular weight and isolelectric point were 110,000 (5.1 S) and 5.0, respectively.

The similarity of enzymatic properties of both the present enzymes to those of other plant carboxypeptidases and pig kidney cathepsin A are discussed.     

Key Odorants in Cured Madagascar Vanilla Beans (Vanilla planiforia) of Differing Bean Quality

The effects of pressure on the enzyme activities of Lacto-bacillus helveticus LHE-511 were examined. Treatment at 400 MPa at 30°C for 10 min completely inhibited acid-producing activity, but the activities of aminopeptidase (AP) and X-prolyl dipeptidyl aminopeptidase (X-PDAP), which are important for acceleration of cheese ripening, increased. These results showed that pressure treatment of L. helveticus LHE-511 selectively inactivates its acid-producing activity.     

Biotransformation of eugenol to vanillin by a novel strain Bacillus safensis SMS1003

Due to the extensive applications of vanillin as flavored compound and increasing consumers concern for its natural and environment friendly mode of production, present work was focused on the selection of bacterial isolate capable of producing vanillin using eugenol biotransformation. Bacterial strain SMS1003 is evidenced as the potential strain for vanillin production and identified as Bacillus safensis (GeneBank accession no. MG561863) using biochemical tests and molecular phylogenic analysis of its 16S rDNA gene sequence. Molar yield of vanillin reached up to 10.7% (0.055?g/L) at 96?h of biotransformation using growing culture of B. safensis SMS1003 in following culture conditions: eugenol concentration 500?mg/L; temperature 37?°C; initial pH 7.0; inoculum volume 4%; volume of culture media 10%; and shaking speed 180?rpm. Vanillin was detected as the single metabolite with a molar yield of 26% (0.12?g/L) at 96?h using resting cells of B. safensis SMS1003. Product confirmation was based on spectral scan using photodiode array detector, Fourier-transform infrared spectroscopy, high-performance liquid chromatography, and mass spectroscopy.     

Efficacy and repellency of some essential oils and their blends against larval and adult house flies,Musca domestica L. (Diptera: Muscidae)

House flies are global pests and notoriously difficult to control. Essential oils of vetiver, cinnamon, and lavender and their blends were tested for toxic and repellent effects against larval and adult flies. All of the oils had moderate toxicity for eggs. Mortality of 2^nd instar larvae was 57–78% in dipping assays, 38–100% in contact assays, and 94–100% in treated media. Lavender was less effective (38% mortality) than the others (91–100%) in contact bioassays. Oil blends were not more effective against larvae than individual oils. Vetiver and cinnamon oils were strongly repellent (84 and 78%, respectively) for larvae in treated media. None of the oils were repellent for adult house flies in olfactometer assays, but testing of additional products demonstrated significant repellency for neem oil, p‐menthane‐3,8‐diol (PMD), and vanillin. Contact/fumigant toxicity of vetiver, cinnamon, and lavender oils was 100%, significantly higher than mortality from sunflower oil (67%). Blends of oils were not more effective against adults than the individual oils, but blends diluted with sunflower oil were as effective as the individual oils. Essentials oils of vetiver and cinnamon may have potential for fly management in situations where conventional insecticides cannot be used.     

吸附剂及培养基组成对香荚兰细胞悬浮培养产生香兰素的影响

本文报道了培养基的组成及添加吸附剂对香荚兰细胞悬浮培养产生香兰素的影响。结果表明,添加吸附剂活性炭及XAD-2后,香荚兰细胞产生的香兰素含量明显增加,而且活性炭的效果优于XAD-2;香荚兰细胞在全组成的MS培养基中香兰素含量均低于由矿物质盐组成培养基中的含量。可以考虑采用二步培养法来培养香荚兰细胞及产生香兰素。     

毛栓菌漆酶诱导及部分特性研究

研究了碳源、氮源、愈创木酚、香兰素及培养条件对漆酶分泌的影响 ;结果表明 ,麦草粉作碳源、(NH4 ) 2 SO4 作氮源有利于漆酶的分泌 ,适宜浓度的愈创木酚和香兰素等对漆酶的产生有一定的作用 ;pH在 3 0 - 8 0的范围内对漆酶的分泌影响差别不大 ,培养温度、接种量、通气量对漆酶的分泌有较大影响。漆酶最适pH值为 4 0 ,最适反应温度为 30℃ ,K+ 、Zn2 + 、Cu2 + 离子可激活漆酶 ;而Ag+ 、Fe3+ 、Cl- 离子可抑制漆酶活性。漆酶的Km值为 1 81× 10 - 3mol L。     

香荚兰细胞悬浮培养产生香兰素条件的研究

该文报道了碳源、氮源及吸附剂对香荚兰细胞悬浮培养产生香兰素的影响,结果表明,蔗糖比葡萄糖及果糖更适合作香荚兰细胞生长及产生香兰素的碳源,最佳蔗糖浓度为５％;当培养基中仅含ＫＮＯ３,则有利于细胞的生长和香兰素的形成,培养液中去掉ＫＮＯ３仅含ＮＨ４ＮＯ３时,细胞生长和香兰素形成均被抑制;培养基添加吸附剂后,香美兰细胞产生的香兰素含量明显增加,活性炭的效果优于ＸＡＤ-２,而且活性炭用量增加,香兰素的产量亦增加。     

Vanillin production from simple phenols by wine-associated lactic acid bacteria

AIMS: The ability of lactic acid bacteria (LAB) to metabolize certain phenolic precursors to vanillin was investigated. METHODS AND RESULTS: Gas chromatography-mass spectrometry (GC-MS) or HPLC was used to evaluate the biosynthesis of vanillin from simple phenolic precursors. LAB were not able to form vanillin from eugenol, isoeugenol or vanillic acid. However Oenococcus oeni or Lactobacillus sp. could convert ferulic acid to vanillin, but in low yield. Only Lactobacillus sp. or Pediococcus sp. strains were able to produce significant quantities of 4-vinylguaiacol from ferulic acid. Moreover, LAB reduced vanillin to the corresponding vanillyl alcohol. CONCLUSIONS: The transformation of phenolic compounds tested by LAB could not explain the concentrations of vanillin observed during LAB growth in contact with wood. SIGNIFICANCE AND IMPACT OF THE STUDY: Important details of the role of LAB in the conversion of phenolic compounds to vanillin have been elucidated. These findings contribute to the understanding of malolactic fermentation in the production of aroma compounds.