Production of Vinegar Mango Using Acetobacter tropicalis CRSBAN-BVA1 and CRSBAN-BVK2 Isolated from Burkina Faso

Production and quality of vinegar from mango juice was evaluated using a two steps production procedure. The first step of fermentation was done using Saccharomyces cerevisae KVL013 for 7 days. The second step, an acetic fermentation was realized using two acetic acid bacteria: A. tropicalis CRSBAN-BVA1 and A. tropicalis CRSBAN-BVK2 for 21 days. Several parameters of the vinegar produced such as physico-chemical and sensory properties were determined. Microbial density during each step was monitored. Results showed that pH, alcoholic and acetic acid contents of vinegar were respectively 2.97%, 7% and 4.54% g/ml respectively by using A. tropicalis CRSBAN-BVA1 and 3.02%, 7% and 4.32% g/ml with A. tropicalis CRSBAN-BVK2. Sensory evaluation revealed that the vinegar was acceptable to the panellists. Results of microbial density showed that the maximum concentration of cell biomass produced was 4.32 × 108 and 4.25 × 108 CFU/ml respectively for CRSBAN-BVA1 and CRSBAN-BVK2.

高浓度苯酚降解菌Candida tropicalis Z-04的鉴定及其对苯酚降解条件的优化

从降解苯酚的好氧颗粒污泥中筛选出一株苯酚降解菌,该菌株最高可耐受2.0 g/L的苯酚。依据形态特征和内源转录间隔区(ITS)的核酸序列分析,将该菌株鉴定为热带假丝酵母菌(Candida tropicalis),并命名为Candida tropicalis Z-04。采用中心复合设计(CCD)法对Candida tropicalis Z-04降解苯酚的条件进行合理设计,利用Design-Expert软件对试验数据进行分析,考察了各因素及其交互作用对苯酚降解率的影响,并得到苯酚最佳降解条件。结果表明,Candida tropicalis Z-04降解苯酚的最佳条件为:酵母提取物投加量0.41 g/L,苯酚初始质量浓度1.03 g/L,菌种投加量1.43%(体积分数),温度30.04℃。在此条件下苯酚的理论预测降解率可达到最大值99.10%。

用基因组改组技术改良发酵木糖酵母Candida tropicalis XY-19的耐乙醇性能

Candida tropicalis XY-19是一株具有优良乙醇发酵性能的发酵木糖酵母,其发酵葡萄糖产乙醇性能与目前酒精工业生产菌种--安琪酒精酵母相近,但XY-19的耐乙醇性能远比安琪酒精酵母差,XY-19在含有超过7%(v/v)乙醇的培养基中不能生长。以XY-19为出发菌株,经紫外线诱变获得了5株能在7.5%(v/v)乙醇的培养基中旺盛生长的突变株,经Co-60诱变获得了8株能在含8%(v/v)乙醇的培养基中旺盛生长的突变株。然后,以紫外线诱变得到的5株菌和Co-60诱变得到的8株菌及耐乙醇性能较好的酿酒酵母(S.cerevisae Angel,S.cerevisae4608和S.cerevisae172)为出发菌株,经过4轮Genome shuffling结合木糖乙醇梯度平板的筛选,获得了4株(G3-13,G3-18,G3-57和G3-60)能够在12%乙醇平板上生长的菌株,其乙醇耐受性比野生菌株XY-19提高了71%,为将XY-19进一步开发成纤维质乙醇发酵的生产菌种奠定基础。本研究结果进一步体现了Genome shuffling技术在改良如乙醇耐受性等多基因控制性状上的突出优势,为工业生产菌种的快速有效改良提供了一种有效的方法。

胺基化磁性壳聚糖微球固定化Candida tropicalis处理含酚废水研究

通过戊二醛将Candida tropicalis固定在胺基化磁性壳聚糖微球（NH2-M-CSm）上，探讨了游离Ct、磁性壳聚糖微球（M-CSm）固定化Ct和NH2-M-CSm固定化Ct除酚的最适条件。结果表明，游离Ct最适pH为5.5，M-CSm和NH2-M-CSm固定化Ct均在4-6之间；游离Ct最适温度为35℃，M-CSm和NH2-M-CSm固定化Ct均为30℃；NH2-M-CSm固定化Ct的热稳定性〉M-CSm固定化Ct〉游离Ct；离子强度对固定化Ct无影响，而对游离Ct有一定的影响；一价态离子对游离Ct和固定化Ct影响不大；二价态离子对游离Ct起抑制作用，而对固定化Ct起促进作用；三价态离子对游离Ct和固定化Ct均起抑制作用。通过实验还得知，固定化Ct可以通过再培养恢复酶活力，其半衰期至少为90 d。最后，在实际废水处理中，经3 h反应后，NH2-M-CSm固定化Ct除酚几乎达100%，而游离Ct只达到80%左右。

葡萄糖-6-磷酸脱氢酶基因的Candida tropicalis过量表达及其对木糖醇合成代谢的影响

研究葡萄糖-6-磷酸脱氢酶基因g6pd过量表达对Candida tropicalis木糖醇生物合成代谢的影响。克隆Candida tropicalis CT16的g6pd基因,并将其与表达载体pYES-pgk重组连接,构建重组载体pYES-pgk-g6pd,LiAc/ssDNA/PEG方法转化导入C.tropicalis CT16,筛选阳性转化子,实现g6pd基因的过量表达。结果表明:发酵62 h,阳性转化子C.tropicalis SYG5的葡萄糖-6-磷酸脱氢酶活力提高了300%,发酵液中木糖醇质量浓度达到79.90 g/L,较野生型对照菌株的木糖醇产量提高了12.41%,木糖醇产率提高了44.94%。因此,葡萄糖-6-磷酸脱氢酶在C.tropicalis木糖醇的合成代谢途径中发挥重要作用,增强g6pd基因的表达,可以明显提高菌体NADPH供应量和还原力,有利于木糖醇的生物合成。

Candida tropicalis infection in a term neonate with gall bladder masses and infective endocarditis

Candida endocarditis is extremely rare in term neonates,and gall bladder involvement due to candidemia has never been reported amongst neonates and infants.A term,appropriate for gestational age neonate developed Candida tropicalis blood stream infection in second week of life.He was started on conventional amphotericin B.However,he failed to show any clinical improvement,and candidemia keep on persisting.Repeat sanctuary sites screening revealed multiple echogenic masses in heart(vegetations) and gall bladder.On changing the treatment to liposomal amphotericin B and fluconazole,he recovered clinically,echogenic masses in gall bladder disappeared,and intracardiac vegetations decreased in size.

Preliminary Characterization of Xylose Reductase Partially Purified by Reversed Micelles from <i>Candida tropicalis</i>IEC5-ITV, an Indigenous Xylitol-Producing Strain

Xylose reductase (EC 1.1.1.21) of Candida tropicalis IEC5-ITV, an indigenous xylitol-producing strain, was partially purified by reversed micelles and characterized, an 8.1 fold purification factor being obtained. The XR present in the crude extract exhibited its highest specific activity at pH 6.0 and 40℃, while in that obtained by reverse micelles, this occurs at pH 6.0 and 30℃. XR before and after extraction is stable within a range of 30 to 40℃, pH 7 after one hour of incubation under these conditions. After two months’storage at –18℃, the enzyme obtained by reverse micelles lost 76.60% specific activity. The estimated molecular weight by PAGE-SDS was 32.42 kD. KM for xylose was higher for the XR extracted by reverse micelles (0.026 M) than that obtained for the enzyme before extraction (0.0059 M), while KM for cofactor NADPH was lower after than before extraction (1.85 mM to 12.0 mM respectively). There was no activity with NADH as a cofactor. Variations in pH and temperature optima, as well as kinetic parameters before and after partial XR purification by reverse micelles are probably due to an alteration in enzyme molecule structure caused by the solvents used during extraction.

Optimization of Candida tropicalis Fermentation Conditions to Produce Long-chain Dibasic Acids

[Objectives] The fermentation conditions of Candida tropicalis 1798-pxa1 were optimized to further improve its yield of long-chain dibasic acids. [Methods] The strains used in the study were C. tropicalis 1798, C. tropicalis 1798-pxa1 and C. tropicalis 1798-pxa1 p2. First, through single factor experiments, the activated three fungi were cultured in different carbon sources, and the absorbance was measured every 2 h. The growth curve was drawn by software, and finally the most suitable substance for the substrate was selected, i.e., dodecane. Then, the composition of the fermentation medium and the fermentation process parameters were determined through the PB experiment of the single-deletion C. tropicalis 1798-pxa1. [Results] The experiment determined the important components affecting the synthesis of long-chain dibasic acids, namely(NH_4)_2SO_4, NaCl and dodecane. After optimization of the culture conditions, the yield of long-chain dibasic acids of C. tropicalis 1798-pxa1 increased from 5.6 to 10.1 g/L. [Conclusions] The scheme has been verified to be capable of greatly increasing the yield of the corresponding long-chain dibasic acids of the C. tropicalis engineering strain.

Model-Based Analysis of a Phenol Bio-Oxidation Process by Adhered and Suspended Candida tropicalis

Phenol is an important commodity for the chemical industry, used for many processes and deemed to be a major pollutant due its xenobiotic nature and high toxicity. For the purpose of phenol bioremediation a biotechnological set up consisting of a continuous packed column bioreactor with Candida tropicalis adhered onto activated carbon beads has been previously described. In this work, we show how the integration of available experimental data of such a biotechnological set up into a mathematical model, can lead both to a better comprehension of the underlying physiological mechanisms operating in the cell culture, and to the identification of the system parameters optimum performance. The model so constructed describes the dynamics of phenol uptake and growth rates by the adhered and suspended biomass;the lethality rates;the adhered biomass removal into suspension or adherence onto carbon beads rates and the phenol and biomass (adhered and suspended) concentrations. It also serves to identify different physiological states for the adhered and the suspended biomass;its predictions being verified by comparing with experimental observations. Based on the model description, different optimization strategies are proposed, some of which have been experimentally tested, encompassing changes in bioreactor operation conditions, process development and strain development.

Determination of <i>α</i>-1,3-Linked Mannose Residue in the Cell Wall Mannan of <i>Candida tropicalis</i>NBRC 1400 Strain

To investigate the chemical structure of cell wall mannan obtained from pathogenic yeast, Candida tropicalis NBRC 1400 (former antigenic standard strain, IFO 1400). As a result of two-dimensional NMR analysis, it was shown that the mannan of this strain is composed of α-1,6-, α-1,3-, α-1,2- and β-1,2-linked mannose residues. In this research, the mannan was subjected to three degradation procedures, acid-treatment, α-mannosidase, and acetolysis under two conditions in order to determine the chemical structure of the antigenic oligomannosyl side chains in this molecule. The 1H-nuclear magnetic resonance spectra of resultant oligosaccharides, pentaose and hexaose, demonstrated the existence of the oligomannosyl side chains corresponding to Manα1-3Manα1-2Manα1-2Manα1-2Man and Manα1-3Manα1-2Manα1-2Manα1-2Manα1-2Man, respectively, which have previously also been found in Candida albicans serotype A strain mannans. These findings indicate that C. tropicalis and C. albicans serotype A have no significant difference in the chemical structure of these cell wall mannans. Therefore, it can be interpreted that it is extremely difficult to distinguish both species by targeting the antigenic group in these mannans.

Antifungal Activity of Oleoresin and Fractions of Pinus elliottii Engelm and Pinus tropicalis against Phytopathogens

Microorganism resistance to the existing products is yet another difficulty that agriculturalists have to deal with. In this context, the search for new agricultural products that can fight phytopathogens has become increasingly important. Plants have played an important role in this process, because they can serve as a source of new compounds for drug discovery. Plants belonging to the genus Pinus produce an oleoresin that protects the plant against herbivores and pathogens. With a view to developing products that can combat fungal pathogens without harming the environment, this work aimed to evaluate the antifungal activity of the oleoresins and fractions of Pinus elliottii Engelm and Pinus tropicalis against phytopathogens. The methodology based on NCCLS M38-A standards aided antifungal activity assessment. The microdilution method helped to determine the Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (MFC). The oleoresins of P. elliottii and P. tropicalis afforded the most significant results—they displayed fungicidal activity against all the tested species. MIC values were promising, especially the MIC of the oleoresin of P. elliottii against S. rolfsii (1.95 μg·mL-1). The MIC values of the oleoresins of P. elliottii and P. tropicalis ranged from 1.95 to 1000 μg·mL-1 and from 31.25 to 250 μg·mL-1, respectively. Fraction PT2 of P. tropicalis furnished the best results among all the assayed fractions: MIC values lay between 125 and 500 μg·mL-11. In conclusion, the oleoresin of P. tropicalis is a promising source of new antifungal agents for application in the treatment of phytopathogenic infections.

Toxic effects of long-term dual or single exposure to oxytetracycline and arsenic on Xenopus tropicalis living in duck wastewater

Direct discharge of aquaculture wastewater may have toxic effects,due to the presence of heavy metals,antibiotics,and even resistant pathogens,but little attention has been given.Here,tanks simulating a wild ecosystem were built to study the effects of long-term exposure to duckwastewater containing oxytetracycline(OTC)and/or arsenic(As)on the growth,physiological function,and gut microbiota evolution of Xenopus tropicalis.The results showed that duck wastewater had no apparent impact on X.tropicalis,but the impact increased significantly(P<0.05)with exposure to OTC and/or As,especially the impact on body weight and growth rate.Biochemical indicators revealed varying degrees of oxidative stress damage,hepatotoxicity(inflammation,necrosis,and sinusoids),and collagen fibrosis of X.tropicalis in all treated groups after 72 days of exposure,which indirectly inhibited X.tropicalis growth.Moreover,16S rDNA amplicon sequencing results showed that the gut microbiota structure and metabolic function were perturbed after chronic exposure,which might be the leading cause of growth inhibition.Interestingly,the abundance of intestinal resistance genes(RGs)increased with exposure time owing to the combined direct and indirect effects of stress factors in duck wastewater.Moreover,once the RGs were expressed,the resistance persisted for at least 24 days,especially that conferred by tetA.These results provide evidence of the toxic effects of DW containing OTC(0.1–4.0 mg/L)and/or As(0.3–3.5μg/L)on amphibians and indicate that it is vital to limit the usage of heavy metals and antibiotics on farms to control the biotoxicity of wastewater.

Optimization of phenol degradation by Candida tropicalis Z-04 using Plackett-Burman design and response surface methodology

Statistical experimental designs were used to optimize the process of phenol degradation by Candida tropicalis Z-04, isolated from phenol-degrading aerobic granules. The most important factors influencing phenol degradation （p 〈 0.05）, as identified by a two-level Plackett-Burman design with 11 variables, were yeast extract, phenol, inoculum size, and temperature. Steepest ascent method was undertaken to determine the optimal regions of these four significant factors. Central composite design （CCD） and response surface analysis were adopted to further investigate the mutual interactions between these variables and to identify their optimal values that would generate maximum phenol degradation. The analysis results indicated that interactions between yeast extract and temperature, phenol and temperature, inocuhim size and temperature affected the response variable （phenol degradation） significantly. The predicted results showed that the maximum removal efficiency of phenol （99.10%） could be obtained under the optimum conditions of yeast extract 0.41 g/L, phenol 1.03 g/L, inoculum size 1.43% （V/V） and temperature 30.04℃. These predicted values were further verified by validation experiments. The excellent correlation between predicted and experimental values confirmed the validity and practicability of this statistical optimum strategy. This study indicated the excellent ability of C. tropicalis Z-04 in degrading high-strength phenol. Optimal conditions obtained in this experiment laid a solid foundation for further use of this microorganism in the treatment of highstrength phenol effluents.

Biodegradation of aniline by Candida tropicalis AN1 isolated from aerobic granular sludge

Aniline-degrading microbes were cultivated and acclimated with the initial activated sludge collected from a chemical wastewater treatment plant. During the acclimation processes, aerobic granular sludge being able to effectively degrade aniline was successfully formed, from which a preponderant bacterial strain was isolated and named as AN1. Effects of factors including pH, temperature, and second carbon/nitrogen source on the biodegradation of aniline were investigated. Results showed that the optimal conditions for the biodegradation of aniline by the strain AN1 were at pH 7.0 and 28–35°C. At the optimal pH and temperature, the biodegradation rate of aniline could reach as high as 17.8 mg/（L·hr） when the initial aniline concentration was 400 mg/L. Further studies revealed that the addition of 1 g/L glucose or ammonium chloride as a second carbon or nitrogen source could slightly enhance the biodegradation efficiency from 93.0% to 95.1%–98.5%. However, even more addition of glucose or ammonium could not further enhance the biodegradation process but delayed the biodegradation of aniline by the strain AN1. Based on morphological and physiological characteristics as well as the phylogenetic analysis of 26S rDNA sequences, the strain AN1 was identified as Candida tropicalis.

Polymorphism analysis of virulence-related genes among Candida tropicalis isolates

Background: Adhesion, biofilm formation, yeast-hyphal transition, secretion of enzymes, and hemolytic activity are all considered important factors in Candida tropicalis infection. However, DNA sequence data for this pathogen are limited. In this study, the polymorphism and heterogeneity of genes agglutinin-like sequences (ALS)2, Lipase (LIP)1, LIP4, and secretory aspartyl proteinase tropicalis (SAPT)1-4 as well as the relationship between phenotype and genotype were analyzed. Methods: This study started in August 2013, and ended in July 2017. The complete length of ALS2, LIP1, LIP4, and SAPT1-4 of 68 clinical C. tropicalis isolates was sequenced. Single nucleotide polymorphisms (SNPs) as well as insertions and deletions (indels) were identified within these genes. In addition, phenotypic characteristics of the virulent factors, including adhesion and the secretion of aspartyl proteinases and phospholipases, were determined. Results: There were 73, 24, 17, 16, 13, and 180 SNPs in the genes LIP1, LIP4, SAPT1, SAPT2, SAPT3, and SAPT4, respectively. Furthermore, 209 SNPs were identified in total for the gene ALS2. Interestingly, large fragment deletions and insertions were also found in ALS2. Isolate FXCT 01 obtained from blood had deletions on all 4 sites and showed the lowest adhesion ability on the polymethylpentene surface. In addition, isolates with deletions in the regions 1697 to 1925 and 2073 to 2272 bp displayed relatively low abilities for adhesion and biofilm formation, and this phenotype correlated with the deletions found in ALS2. LIP1, SAPT4, and ALS2 displayed great heterogeneity among the isolates. Large deletions found in gene ALS2 appeared to be associated with the low ability of adhesion and biofilm formation of C. tropicalis. Conclusion: This study might be useful for deeper explorations of gene function and studying the virulent mechanisms of C. tropicalis.

Histological observation on unique phenotypes of malformation induced in Xenopus tropicalis larvae by tributyltin

Tributyltin（TBT）,a biocide used in antifouling paints,has shown strong teratogenic effects on Xenopus tropicalis embryos at environmentally relevant concentrations.X.tropicalis embryos were exposed to 50,100 and 200 ng/L tributyltin chloride for 72 hr.The histological changes were further observed on abnormal eyes,enlarged trunks,enlarged proctodaeums and absence of fins induced by TBT.The lens and the retinal layers of abnormal eyes were slightly or barely differentiated,and that the pigment epithelium was neither continuous nor smooth.The abdomens were full of undifferentiated gut tissue with yolk-rich inclusions in the tadpoles with enlarged trunks.The proctodaeums formed a bump-like or columnar structure.The mass of yolk-rich cells occupied the lumen,blocked the opening and even turned inside out of the proctodaeum.Both the ventral and dorsal fins in trunks and tails became narrow or even disappeared totally.Our results suggest that great changes of histology took place corresponding to the unique phenotypes.The gut tissue was poorly differentiated,which led to the failed elongation of the guts and subsequently the enlarged trunks.The enlarged proctodaeums were due to the undifferentiation of inner layer,the expansion of outer epidermal part and the absence of fins around them.In brief,the histological observations provided insights into the reason of the unique external malformations in some degree.

Ambient temperature alters body size and gut microbiota of Xenopus tropicalis

Temperature is important to determine physiological status of ectotherms. However, it is still not fully understood how amphibians and their symbiotic microbiota acclimate to ambient temperature. In this study, we investigated the changes of gut microbiota of Xenopus tropicalis at different temperatures under controlled laboratory conditions. The results showed that microbial communities were distinct and shared only a small overlap among froglet guts, culture water and food samples.Furthermore, the dominant taxa harbored in the gut exhibited low relative abundance in water and food. It indicates that bacterial taxa selected by amphibian gut were generally of low abundance in the external environment. Temperature could affect betadiversity of gut microbiota in terms of phylogenetic distance, but it did not affect alpha diversity. The composition of gut microbiota was similar in warm and cool treatments. However, signature taxa in different temperature environments were identified. The relationships between temperature, gut microbiota and morphology traits of X. tropicalis revealed in this study help us to predict the consequences of environmental changes on ectothermic animals.

Multilocus sequence typing indicates diverse origins of invasive Candida tropicalis isolates in China

Background According to data from the China Hospital Invasive Fungal Surveillance Net （CHIF-NET） 2010, Candida tropica/is （C. tropica/is） is the third most common pathogen causing invasive candidiasis. Moreover, the majority of fluconazole-resistant C. tropicalis isolates were from a single hospital. Therefore, a molecular epidemiological survey is necessary to investigate the genetic relatedness of C. tropica/is isolates in China. Methods In this study, 48 C. tropicalis isolates causing invasive fungal infections from four tertiary hospitals in China were studied. All the isolates were identified by sequencing the internal transcribed spacer region. Antifungal susceptibility to triazoles, amphotericin B, and caspofungin was determined by the Clinical and Laboratory Standards Institute standard broth microdilution method. Multilocus sequence typing （MLST） was performed, and phylogenetic analysis was further performed by the eBURST and maximum parsimony （MP） methods to characterize the genetic relatedness of isolates. Results MLST discriminated 40 diploid sequence types （DSTs） among 48 isolates, including 36 novel DSTs, and the XYR1 gene showed the highest discriminatory power. The DSTs obtained from this study were compared with those of previously reported C. tropicalis isolates, and there was poor type alignment with regional strains. Nine groups and 11 singletons were identified by eBURST, whereas two groups and 10 subgroups were clustered by MP analysis. Generally, there were no obvious correlations between clonal clusters generated and the specimen source or hospital origin. Seven fiuconazole-resistant isolates were confirmed and assigned to three distinguishable branches. Conclusions The results suggested diverse origins of invasive C. tropicalis isolates in China. Although most invasive C. tropicalis strains in the mainland of China were clustered with previously characterized Asian isolates, major C. tropicalis clusters identified in this study were genetically distinct from those of other geographic regions.

Unexpected phenotypes of malformations induced in Xenopus tropicalis embryos by combined exposure to triphenyltin and 9-cis-retinoic acid

Xenopus tropicalis embryos were exposed for 48 hr to the mixtures of 5 μg Sn/L triphenyltin （TPT）, which is a well-known endocrine disruptor, and 0.25-5 μg/L 9-cis retinoic acid （9c-RA）, which is the natural ligand of retinoid X receptor. The phenotypes induced by combined exposure were more variable than those resulting from single exposure to either TPT or 9c-RA. The prominent phenotypes included underdeveloped head structures, abnormal eyes, narrow fins, enlarged proctodaeum, etc. Especially, combined exposure induced unexpected notochord malformations, which ranged from small swellings of the surface of the tails to the extension and extrusion of notochord out of the posterior tails. Compared with the 5 μg Sn/L TPT-treated group, the index of fin deficiency was not affected, and the index of axis deficiency was significantly increased with increasing RA concentrations in the mixtures. Our results suggest that combined exposure to TPT and 9c-RA induced not only more variable phenotypes of malformations than exposure to single compound but also some new and unexpected phenotypes.

高浓度有机废水酵母菌低氧处理技术初探

在开放式环境中,在低氧条件(DO=0-0.2mg/L)下通过连续小试试验考察了利用酵母Candidatropicalis处理高浓度有机废水的可行性。54d低氧连续小试结果表明:进水COD、NH4+-N、TP浓度分别为11000～30000、1000、230mg/L时,其去除率分别为67%～88%,18%～68%,23%～68%,氮磷去除率取决于进水COD、TN、TP的水平。污泥微观形态观察表明反应柱内微生物基本上是酵母,但运行过程中优势酵母形态发生了变化。低氧(DO=0.1-0.4mg/L)和无氧条件下C.tropicalis批量培养试验表明污染物去除和酵母生长与氧的供给关系密切,低氧条件下C.tropicalis有很高的活性,能有效去除COD(去除率为91%);无氧条件则导致C.tropicalis生物量下降,COD去除率仅为12%。