Candida molischianaβ-Glucosidase Production by Saccharomyces cerevisiae and its Application in Winemaking

ABSTRACT: A recombinant Saccharomyces cerevisiae wine yeast strain expressing the Candida molischiana bgln gene encoding a β-glucosidase (BGLN) has been used to produce this enzyme. Shaking rate, pH, and aeration rate conditions have been optimized to obtain maximum activity to facilitate enzyme purification. The ability of the heterologous enzyme to efficiently release terpenols and alcohols from a Muscat wine glycoside extract and also directly from wine has been demonstrated. Terpenol glycoside content decreased by 50% after 1 mo of wine storage in agreement with results reported for the β-glucosidase produced by C. molischiana.     

Modeling for local dynamic behaviors of phenol biodegradation in bubble columns

A coupled three-dimensional (3-D) model, combining hydrodynamics with biochemical reactions, was developed to simulate the local transient flow patterns and the dynamic behaviors of cell growth and phenol biodegradation by yeast Candida tropicalis in the bubble-column bioreactor, using the computational fluid dynamic (CFD) method. In order to validate this proposed model effectively, the validation of the local hydrodynamic characteristics of the gas-mineral salt solution (gas-liquid) two-phase system, with the phenol concentration of 1200 mg/L, and with the absence of cells, was performed in a square-sectioned bubble column bioreactor using the LDA system and conductivity probe. Furthermore, the validation of phenol biodegradation behaviors by yeast Candida tropicalis at different initial concentrations of phenol and cell was also carried out in the above bubble-column bioreactor. The results indicated that the model simulations had a satisfying agreement with the experimental data. Finally, the local instantaneous flow and phenol biodegradation features including gas holdup, gas velocity, liquid velocity, cell concentration and phenol concentration inside the bioreactor were successfully predicted in different-scale bubble columns by the proposed model. © 2006 American Institute of Chemical Engineers AIChE J, 2006     

Concentration of docosahexaenoic acid in glyceride by hydrolysis of fish oil withcandida cylindracea lipase

In an attempt to concentrate the content of DHA (docosahexaenoic acid) in a glyceride mixture containing triglyceride, diglyceride and monoglyceride, fish oil was hydrolyzed with six kinds of microbial lipase. After the hydrolysis, free fatty acid was removed and fatty acid components of the glyceride mixtures were analyzed. When the hydrolysis withCandida cylindracea lipase was 70% complete, the DHA content in the glyceride mixture was three times more than that in the original fish oil. The EPA (eicosapentaenoic acid) content became almost 70% of the original fish oil. Hydrolysis with other lipases did not result in an increase in the DHA content in the glyceride mixtures. Hydrolysis of DHA-rich tuna oil (DHA content is about 25%) withCandida cylindracea lipase resulted in 53% DHA in the glyceride mixture. The EPA content, however, remained close to that of the original tuna oil. In this report, the acyl chain specificity of lipases is evaluated in terms of hydrolysis resistant value (HRV). HRV is the ratio between the DHA contents in the glyceride mixture of hydrolyzed oil and original oil. HRV clearly indicates differences in hydrolysis between DHA and other fatty acids (e.g., saturated and monoenoic acids).     

Assignment of most genes encoding major peroxisomal polypeptides to chromosomal band V of the asporogenic yeast Candida tropicalis

The peroxisomes of the asporogenic yeast Candida tropicalis contain about 20 major polypeptides (PXPs). We have isolated a number of genes encoding them; 11 POX genes encoded independent PXPs and three POY genes were likely to encode three other PXPs. To locate these genes on the chromosomes, chromosomes of C. tropicalis were separated by pulsed-field gel electrophoresis. Eight chromosomal bands were observed over the range of 1.0 Mbp (band 1) to 2.8 Mbp (band VIII); the genome size was estimated to be about 20 Mbp. Southern blot analysis showed that ten genes were on band V, three genes were on band IV, and the other gene was on band VI. Three genes gave hybridization signals of nearly equal intensity on two different chromosomal bands: POX6A and POX8B, on bands V and VII; and POX8A, on bands IV and VI. Ribosomal RNA genes also hybridized to two bands, VI and VII. Most genes assigned to only one band hybridized to two restriction fragments produced by either NotI or SfiI endonuclease. The results suggested that C. tropicalis was diploid and that restriction sites were conserved little between homologues. The three POX genes that were found on two chromosomal bands hybridized to not more than two restriction fragments, implying that the allelic genes were present on different chromosomal bands.     

Interplay between Humoral and CLA+ T Cell Response against Candida albicans in Psoriasis

Candida albicans (CA) infections have been associated with psoriasis onset or disease flares. However, the integrated immune response against this fungus is still poorly characterized in psoriasis. We studied specific immunoglobulins in plasma and the CA response in cocultures of circulating memory CD45RA⁻ cutaneous lymphocyte antigen (CLA)^+/− T cell with autologous epidermal cells from plaque and guttate psoriasis patients (cohort 1, n = 52), and also healthy individuals (n = 17). A complete proteomic profile was also evaluated in plaque psoriasis patients (cohort 2, n = 114) regarding their anti-CA IgA levels. Increased anti-CA IgA and IgG levels are present in the plasma from plaque but not guttate psoriasis compared to healthy controls. CA cellular response is confined to CLA⁺ T cells and is primarily Th17. The levels of anti-CA IgA are directly associated with CLA⁺ Th17 response in plaque psoriasis. Proteomic analysis revealed distinct profiles in psoriasis patients with high anti-CA IgA. C-C motif chemokine ligand 18, chitinase-3-like protein 1 and azurocidin were significantly elevated in the plasma from plaque psoriasis patients with high anti-CA levels and severe disease. Our results indicate a mechanism by which Candida albicans exposure can trigger a clinically relevant IL-17 response in psoriasis. Assessing anti-CA IgA levels may be useful in order to evaluate chronic psoriasis patients.     

响应面优化褶皱假丝酵母脂肪酶催化合成木质甾醇油酸酯

以木质甾醇转化率为指标,考察了10种常见商业化脂肪酶催化合成木质甾醇油酸酯的效果,确定褶皱假丝酵母脂肪酶(CRL)为优选生物催化剂,进一步筛选出正己烷为优选反应介质.在脂肪酶用量、油酸和木质甾醇的物质的量比、反应温度和反应时间这4个单因素考察基础上,通过响应面分析法对酶催化木质甾醇油酸酯合成工艺条件进行优化,并对优化条件进行验证和放大实验.CRL催化合成木质甾醇油酸酯的优化工艺参数为:CRL添加量为木质甾醇质量的10％,油酸与木质甾醇的物质的量比为3.8:1,反应温度为46℃,反应时间为28 h,木质甾醇的转化率为91.56％±0.25％.     

Enzymatic modification of trilinolein: Incorporation of n-3 polyunsaturated fatty acids

Two immobilized lipases, IM60 fromMucor miehei and SP435 fromCandida antarctica, were used as biocatalysts for the modification of trilinolein with n-3 polyunsaturated fatty acids (PUFA), such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), by using their ethyl esters as acyl donors (EEPA and EDHA, respectively). Transesterification (ester-ester interchange) reactions were carried out in organic solvent. The products were analyzed according to their equivalent carbon number and polarity by reverse-phase high-performance liquid chromatography, and the fatty acid profiles were determined by gas-liquid chromatography. Modified triacylglycerol products contained 1 or 2 molecules of n-3 PUFA. With EEPA as the acyl donor, the total EPA product yields with IM60 and SP435 as biocatalysts were 79.6 and 81.4%, respectively. However, with EDHA as the acyl donor and IM60 and SP435 as biocatalysts, the total DHA product yields were 70.5 and 79.7%, respectively. Effects of reaction parameters, such as type of solvent, enzyme load, time course, and molar ratio of substrates on the n-3 PUFA incorporation, were followed with SP435 as the biocatalyst. High yields were obtained, even in the absence of organic solvent. These lipids do hold promise for specialty nutrition and other therapeutic uses.