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.

Protective effect of long-chain polyunsaturated fatty acids on hepatorenal syndrome in rats

BACKGROUND Hepatorenal syndrome(HRS)is the most prevalent form of acute kidney injury in cirrhotic patients.It is characterized by reduced renal blood flow and represents the most severe complication in cirrhotic patients with advanced disease.Previous research has indicated that antioxidants can delay the onset of a hyperdynamic circulatory state in cirrhosis and improve renal function in HRS patients.Regular omega-3 supplementation has significantly reduced the risk of liver disease.This supplementation could represent an additional therapy for individuals with HRS.AIM To evaluated the antioxidant effect of omega-3 polyunsaturated fatty acid supplementation on the kidneys of cirrhotic rats.METHODS Secondary biliary cirrhosis was induced in rats by biliary duct ligation(BDL)for 28 d.We used 24 male Wistar rats divided into the following groups:I(control);II(treated with omega-3,1 g/kg of body weight);III(BDL treated with omega-3,1 g/kg of body weight);and IV(BDL without treatment).The animals were killed by overdose of anesthetic;the kidneys were dissected,removed,frozen in liquid nitrogen,and stored in a freezer at-80℃for later analysis.We evaluated oxidative stress,nitric oxide(NO)metabolites,DNA damage by the comet assay,cell viability test,and apoptosis in the kidneys.Data were analyzed by one-way analysis of variance,and means were compared using the Tukey test,with P≤0.05.RESULTS Omega-3 significantly decreased the production of reactive oxygen species(P<0.001)and lipoperoxidation in the kidneys of cirrhotic rats treated with omega-3(P<0.001).The activity of the antioxidant enzymes superoxide dismutase and catalase increased in the BDL+omega-3 group compared to the BDL group(P<0.01).NO production,DNA damage,and caspase-9 cleavage decreased significantly in the omega-3-treated BDL group.There was an increase in mitochondrial electrochemical potential(P<0.001)in BDL treated with omega-3 compared to BDL.No changes in the cell survival index in HRS with omega-3 compared to the control group(P>0.05)were observed.CONCLUSION The study demonstrates that omega-3 can protect cellular integrity and function by increasing antioxidant enzymes,inhibiting the formation of free radicals,and reducing apoptosis.

Long-chain acyl-CoA synthetase in fatty acid metabolism involved in liver and other diseases:An update

Long-chain acyl-Co A synthetase(ACSL) family members include five different ACSL isoforms, each encoded by a separate gene and have multiple spliced variants. ACSLs on endoplasmic reticulum and mitochondrial outer membrance catalyze fatty acids with chain lengths from 12 to 20 carbon atoms to form acyl-Co As, which are lipid metabolic intermediates and involved in fatty acid metabolism, membrane modifications and various physiological processes. Gain- or lossof-function studies have shown that the expression of individual ACSL isoforms can alter the distribution and amount of intracellular fatty acids. Changes in the types and amounts of fatty acids, in turn, can alter the expression of intracellular ACSLs. ACSL family members affect not only the proliferation of normal cells, but the proliferation of malignant tumor cells. They also regulate cell apoptosis through different signaling pathways and molecular mechanisms. ACSL members have individual functions in fatty acid metabolism in different types of cells depending on substrate preferences, subcellular location and tissue specificity, thus contributing to liver diseases and metabolic diseases, such as fatty liver disease, obesity, atherosclerosis and diabetes. They are also linked to neurological disorders and other diseases. However, the mechanisms are unclear. This review addresses new findings in the classification and properties of ACSLs and the fatty acid metabolismassociated effects of ACSLs in diseases.

Impacts of the unsaturation degree of long-chain fatty acids on the volatile fatty acid profiles of rumen microbial fermentation in goats in vitro

This study investigated the impacts of the degree of unsaturation （unsaturity） of long-chain fatty acids on volatile fatty acid （VFA） profiles of rumen fermentation in vitro. Six types of long-chain fatty acids, including stearic acid （C18：0, control group）, oleic acid （C18：1, n-9）, linoleic acid （C18：2, n-6）, a-linolenic acid （C18：3, n-3）, arachidonic acid （C20：4, n-6） and eicosapentaenoic acid （C20：5, n-3）, were tested. Rumen fluid from three goats fitted with ruminal fistulae was used as inoculum and the inclusion rate of long-chain fatty acid was at 3% （w/w） of substrate. Samples were taken for VFA analysis at 0, 3, 6, 9, 12, 18 and 24 h of incubation, respectively. The analysis showed that there were significant differences in the total VFA among treatments, sampling time points, and treatment×time point interactions （P〈0.01）. a-Linolenic acid had the highest total VFA （P〈0.01） among different long-chain fatty acids tested. The molar proportion of acetate in total VFA significantly differed among treatments （P〈0.01） and sampling time points （P〈0.01）, but not treatment×time point interactions （P〉0.05）. In contrast, the molar proportion of propionate did not differ among treatments during the whole incubation （P〉0.05）. However, for butyrate molar proportions, significant differences were found not only among sampling time points but also among treatments and treatment×time point interactions （P〈0.01）, with eicosapentaenoic acid having the highest value （P〈0.01）. Additionally, no statistically significant differences were found in the acetate to propionate ratios among treatments groups （P〉0.05）, even the treatments stearic acid and a-linolenic acid were numerically higher than the others. The inclusion of 3% long-chain unsaturated fatty acids differing in the degree of unsaturation brought out a significant quadratic regression relation between the total VFA concentration and the double bond number of fatty acid. In conclusion, the a-linolenic acid with 3 double bonds appeared better for improving rumen microbial fermentation and the total VFA concentration.

Roles of Long-chain Acyl Coenzyme A Synthetase in Absorption and Transport of Fatty Acid

Long-chain acyl coenzyme A synthetase(ACSL) is a member of the synthetase family encoded by a multigene family;it plays an important role in the absorption and transport of fatty acid.Here we review the roles of ACSL in the regulating absorption and transport of fatty acid,as well as the connection between ACSL and some metabolic diseases.

Effects of long-chain fatty acid supplementation on the growth performance of grower and finisher pigs: a meta-analysis

Background: Supplementation of feed with long-chain fatty acids(LCFAs) during the grower and finisher phases has long been discussed as a growth promotion strategy in pigs, but its effects are inconsistent. The purpose of our study was to comprehensively evaluate its effects on the growth performance based on the average daily gain(ADG), average daily feed intake(ADFI) and gain: feed(G:F) ratio and to unveil the roles of the basal diet, LCFA concentration and LCFA saturation.Results: We searched the Pub Med and Web of Science databases(articles published from Jan 1 st, 2000, to Sep 30 th,2018;restricted to English) and compared LCFA-supplemented diets with control diets. We retrieved 2346 studies, 18 of which(1314 pigs, 26 records) were eligible for our analysis. We used a random-effects model to calculate the weighted mean differences(WMDs) and 95% confidence intervals(CIs). LCFA supplementation in the grower-finisher phase improved the ADG(WMD = 41.74 g/d, 95% CI: 8.81 to 74.66, P = 0.013) and G:F ratio(WMD = 0.019, 95% CI: 0.006 to 0.032, P = 0.003). For supplementation solely in the finisher phase, we found a similar performance in the ADG(WMD = 39.93 g/d, 95% CI: 26.48 to 53.38, P < 0.001) and G:F ratio(WMD = 0.019, 95% CI: 0.006 to 0.032, P < 0.001) but a reduction in the ADFI(WMD =-83.863 g/d, 95% CI:-156.157 to-11.569, P = 0.023). Specifically, approximately 5%LCFA supplementation in the finisher phase had significant effects on the ADG(WMD = 51.385 g/d, 95% CI: 35.816 to66.954, P < 0.001), ADFI(WMD =-102.869 g/d, 95% CI:-189.236 to-16.502, P = 0.02) and G:F ratio(WMD = 0.028, 95%CI: 0.018 to 0.039, P < 0.001), whereas a concentration of approximately 1% exhibited no effects.Conclusions: Overall, regardless of the basal diet and saturation, LCFA supplementation greatly improves the growth performance of grower and finisher pigs, primarily by increasing the energy density.

Identification of a Long-Chain Fatty Acid Elongase from Nannochloropsis sp. Involved in the Biosynthesis of Fatty Acids by Heterologous Expression in Saccharomyces cerevisiae

The marine microalga Nannochloropsis sp.contains various elongases and desaturases that are critical for biosynthesis of polyunsaturated fatty acids.A full-length cDNA encoding a long-chain fatty acid elongase,named Ns FAE,was cloned from Nannochloropsis sp..The open reading frame of Ns FAE(GenBank accession no.MF680548)consisted of 1068 bp and encoded a predicted protein of 355 amino acids with molecular mass 38.8 k Da.The deduced polypeptide showed 43%–44%identity to fatty acyl elongases from other algae.RT-PCR experiments indicated that the Ns FAE gene exhibited the highest expression in Nannochloropsis sp.at 72 h(i.e.,during the third growth stage)and the expression was significantly lower in the other four growth stages.Plasmid p Ns FAE-CRISPR and a recombinant DNA fragment(ADH1p-Ns FAE-CYCt)were transformed into Saccharomyces cerevisiae strain BY4742 using the CRISPR-Cas system.Yeast transformants containing Ns FAE produced three fatty acids not normally present in wild-type BY4742-linoleic acid,linolenic acid and eicosadienoic acid-indicating that Ns FAE encodes a functional elongase enzyme.

Long-chain alkanol-alkyl carboxylic acid-based low-viscosity hydrophobic deep eutectic solvents for one-pot extraction of anthraquinones from Rhei Radix et Rhizoma

Natural long-chain alkanol and alkyl carboxylic acid were used to prepare novel hydrophobic deep eutectic solvents(HDESs).These HDESs are liquid at room temperature and have low viscosity(<12.26 mPa·s),low polarity(lower than that of methanol,ChCl-based deep eutectic solvents and other reported HDESs),and low density(<0.928 g/mL).A simple one-pot method based on a novel HDES-water two-phase extraction system was constructed for the extraction of weak-polarity bioactive components,anthraquinones,from Rhei Radix et Rhizoma.This HDES-based new extraction method does not consume hazardous organic solvents and can obtain a total anthraquinone yield of 21.52 mg/g,which is close to that obtained by the Chinese pharmacopoeia method(21.22 mg/g)and considerably higher than those by other reported HDESs-based extraction methods(14.20-20.09 mg/g,p<0.01).The high extraction yield can be mainly attributed to the severe destruction of the RRR cell walls by the extraction system and the excellent dissolving ability of novel HDESs for anthraquinones.

The effects of the unsaturated degree of long-chain fatty acids on the rumen microbial protein content and the activities of transaminases and dehydrogenase in vitro

This study investigated the effects of the degree of unsaturation（unsaturity） of long-chain fatty acids on microbial protein content and the activities of transaminases and dehydrogenase in vitro using goat rumen fluid as the cultural medium.Six types of fatty acids,stearic acid（C18：0,group A,control group）,oleic acid（C18：1,n-9,group B）,linoleic acid（C18：2,n-6,group C）,α-linolenic acid（C18：3,n-3,group D）,arachidonic acid（C20：4,n-6,group E）,and eicosapentaenoic acid（C20：5,n-3,group F）,were tested,and the inclusion ratio of each fatty acid was 3%（w/w） in total of culture substrate.Samples were taken at 0,3,6,9,12,18 and 24 h,respectively,during culture for analyses.Compared with stearic acid,linoleic acid,a-linolenic acid,and arachidonic acid increased the bacterial protein content,while oleic acid and eicosapentaenoic acid had no significant effects;the protozoal protein content was reduced for all the unsaturated fatty acids,and the magnitude of the reduction appeared to be associated with the degree of unsaturity of fatty acids.The total microbial protein content was dominantly accounted by the protozoal protein content（about 4-9 folds of the bacterial protein）,and only increased by linoleic acid,but reduced by oleic acid,arachidonic acid and eicosapentaenoic acid.There were no significant effects in the activities of both glutamic oxaloacetic transaminase（GOT） and glutamic-pyruvic transaminase（GPT） for all the other fatty acids,except for arachidonic acid which enhanced GOT activity and oleic acid which enhanced GPT activity.The total dehydrogenase activity was positively correlated with the degree of unsaturation of fatty acids.In conclusion,the inclusion of 3%of long-chain unsaturated fatty acids increased bacterial protein content,whereas reduced protozoal protein content and enhanced dehydrogenase activity.The fatty acids with more than three double bonds had detrimental effects on the microbial protein content.This work demonstrates for the first time the effect rule of the unsaturation degree of long-chain fatty acids on the rumen microbial protein in vitro.

Study on Extracting Rare Earth from Sulfate System by Long-Chain Fatty Acid

The extraction of rare earths by long-chain fatty acid in kerosene from sulphate system was described.It was demonstrated from the experimental results that the ratio of kerosene: fatty acid: isooctanol = 55 : 30: 15 ( V/V),By the saturation capability method and the slope method, the extracted reaction mechanism of the extraction of rare earth was studied.It is shown that the extraction reaction conform to the cation exchange reaction mechanism.The extracted sequence of rare earth was determined in this system and it is shown that there is no tetrad effect and the position of yttrium is between lanthanum and cerium.

POM-ILs的制备及其在聚混合二元酸乙二醇酯合成中的应用

制备了以Sn、Fe、Ti、Zn为配位金属的Keggin型单取代多金属氧酸盐离子液体(POM-ILs)催化剂,利用FTIR、XRD、SEM、EDS及TGA对催化剂进行了表征和测试,将其用于催化混合二元酸二甲酯(DBE)与碳酸乙烯酯(EC)反应合成聚混合二元酸乙二醇酯(PDBE),考察了不同配位金属对POM-ILs催化活性的影响。采用FTIR、GPC、TGA、DSC和微机控制电子万能试验机对PDBE进行了测试,并对PDBE的酶降解性能进行了测试。结果表明,Sn单取代的锌钨酸盐离子液体[Bmim]_(6)ZnW_(11)SnO_(39)(H_(2)O)的催化性能最优,在n(EC)∶n(DBE)=1.5∶1、反应时间5 h、温度215℃、催化剂用量为总原料质量的1.1%的条件下,PDBE预聚物选择性为57.85%,收率为54.76%。在反应时间3.5 h、反应温度220℃的最佳条件下,制备的PDBE特性黏数达到0.591dL/g,拉伸强度为33.4 MPa,断裂伸长率为17.9%;数均相对分子质量为14433,重均相对分子质量为33429,平均相对分子质量为70859,多分散性指数(PDI)为2.3161。当添加质量浓度为1.2 g/L固化脂肪酶435的磷酸盐缓冲溶液降解至质量剩余率为2%~3%时,PDBE需28 d,而聚丁二酸乙二醇酯需30 d。

从混合二元酸中分离戊二酸的研究

从混合二元酸中分离戊二酸是国际上较为热门的课题之一,本文主要对目前常用的一些方法进行了介绍:结晶分离法、萃取结晶法、吸附法、反应精馏法、尿素络合法以及成盐酸化法。但在工业生产中,多种常用的分离技术均无法对混合二元酸进行有效分离,造成戊二酸纯度较低。如果能够从混合二元酸里将戊二酸有效的分离出来,对工业生产而言意义重大。

Natural long-chain saturated fatty acids doped LNPs enabling spleen selective mRNA translation and potent cancer immunotherapy

Rationally tailored lipid nanoparticles(LNPs)with efficient and tunable delivery of mRNA in vivo are crucial for mRNA vaccines.Selective expression of antigenic protein in lymphoid tissues/organs could improve the immunostimulatory efficacy and safety of LNPs-based mRNA vaccines.Inspired by the metabolic behavior that long-chain saturated fatty acids tending to enter lymphoid tissue rather than the liver,we developed fatty acid-doped LNPs capable of mediating differential protein expressions in the liver and spleen when administered intravenously.When the molar ratio of saturated fatty acid located 60%–70%,the doped LNPs achieved the spleen selective mRNA translation.The mechanism could be attributed to the different cellular uptake behaviors of saturated fatty acids in hepatocytes.Immunization with a model antigen(ovalbumin)mRNA-loaded spleen selective LNPs,we observed enhanced antigen-specific T cell immune responses,and potent immunotherapeutic and immunoprophylactic efficacy in the mouse lymphoma model.Our natural long-chain saturated fatty acids metabolic characteristics-inspired design of LNPs for spleen-selective mRNA vaccines delivery will provide references for designing mRNA vaccines with high efficacy and safety for tumor immunotherapy.

高效液相色谱法测定己二酸生产废液中的酸含量

提出了1种采用高效液相色谱法测定己二酸生产废液中硝酸及二元酸含量的方法。通过考察样品稀释倍数的影响,确定样品前处理条件。进行了高效液相色谱仪仪器测试波长及流动相配比的选择试验,确定以体积比2∶8的甲醇∶磷酸缓冲溶液(pH=2.5)为流动相,柱温30℃,测试波长210nm的测试条件,实现了采用高效液相色谱法测定己二酸生产废液中硝酸及二元酸含量。加标回收试验的回收率为96.55%~102.31%,各测试组分相对标准偏差(n=6)均小于5%。方法简便、准确快速,适用于己二酸生产废液中硝酸及二元酸含量的测定。

混合二元酸提取戊二酸工业化研究

基于戊二酸的来源及需求的重要性分析,根据戊二酸与其他2种二元酸的性质差异为理论依据设计了提取戊二酸的工业化工艺流程,同时对该工艺流程的副产物处理进行了分析,该工艺并不产生多余的污染物为绿色化工工艺,因此混合二元酸提取戊二酸工业化具有不仅为企业创造可观的经济效益而且促进化工产业发展的重要意义。

混二元酸二异丁酯的合成及其在建筑内墙涂料中的应用研究

利用已二酸工业生产的副产物混二元酸作为原料,通过采用正交试验对其酯化反应的合成工艺进行优化研究,筛选出最佳工艺条件,制备了一种性能优良的成膜助剂混二元酸二异丁酯。并对该成膜助剂进行了在建筑内墙涂料用乳液及建筑内墙乳胶涂料中应用测试评价,实验结果表明,合成的成膜助剂在建筑内墙涂料中的应用效果和性能与当前市场上普遍使用的十二酯醇成膜助剂完全相当,满足建筑内墙涂料对成膜助剂的使用性能要求。

Fatty acid metabolism and acyl-CoA synthetases in the liver-gut axis

Fatty acids are energy substrates and cell components which participate in regulating signal transduction,transcription factor activity and secretion of bioactive lipid mediators.The acyl-CoA synthetases(ACSs)family containing 26 family members exhibits tissue-specific distribution,distinct fatty acid substrate preferences and diverse biological functions.Increasing evidence indicates that dysregulation of fatty acid metabolism in the liver-gut axis,designated as the bidirectional relationship between the gut,microbiome and liver,is closely associated with a range of human diseases including metabolic disorders,inflammatory disease and carcinoma in the gastrointestinal tract and liver.In this review,we depict the role of ACSs in fatty acid metabolism,possible molecular mechanisms through which they exert functions,and their involvement in hepatocellular and colorectal carcinoma,with particular attention paid to long-chain fatty acids and small-chain fatty acids.Additionally,the liver-gut communication and the liver and gut intersection with the microbiome as well as diseases related to microbiota imbalance in the liver-gut axis are addressed.Moreover,the development of potentially therapeutic small molecules,proteins and compounds targeting ACSs in cancer treatment is summarized.

The determination of acidity of the dilute solutions of weak multibasic organic acids

The new theoretical method for the accurate determination of acidity of dilute solutions of weak multibasic organic acids (which are widely used in medicine, pharmacology, various branches of industry and participate in important biological processes in living organisms) is suggested. The concepts of the contributions of the separate dissociation steps to the [H+] value, xm, are used for an analysis of complex equilibria of the processes of dissociation of these acids. The cases of weak dibasic and tribasic organic acids with the “overlapping” dissociation equilibria and a general case of weak multibasic acids, HnA, are considered. From the conditions of equality of the concentrations of various ionized and non-ionized forms in the dilute solutions of weak multibasic organic acids the areas of dominance of these forms in connection with the corresponding xm values are formulated.

Impact of Chain Length of Saturated Fatty Acids during Their Heterogeneously Catalyzed Deoxygenation

Fatty acids with different chain length were deoxygenated in the absence of hydrogen (caprylic acid (CA), lauric acid (LA) and stearic acid (SA)). The catalytic tests were carried over Pd-containing catalysts out in a batch reactor under inert gas for 6 h at 250&degC to 350&degC and pressures from 18 to 75 bar in the absence of additionally fed hydrogen. Pd-containing catalysts were tested;the best performing catalyst was 10% Pd/C with 63% undecane yield at 327&degC. These catalysts were used for a comparative decarboxylation of CA, LA and SA. At equal reaction conditions (300&degC, 6 h), the chain length of the fatty acid had a strong impact on the conversion, which was steadily increasing, whereas the alkane selectivity ran through a maximum. This work demonstrated the usability of Pd-containing catalysts for the decarboxylation of various fatty acids in the absence of additionally fed hydrogen with respect to the manufacture of hydrocarbons that can be used as blending components for fuels.

混合二元酸二甲酯的合成工艺研究

以混合二元酸(DBA)和甲醇为原料,以对甲苯磺酸为催化剂,不加带水剂的情况下合成混合二元酸二甲酯(DBE),考察不同反应条件对产品收率的影响。最佳工艺条件为:催化剂用量占DBA质量的4%,甲醇与DBA摩尔比为5.6,反应温度110℃,反应时间5 h,此工艺条件下得到的混合二元酸二甲酯收率达到93%。