Evaluation of the intracellular lipid-lowering effect of polyphenols extract from highland barley in HepG2 cells

Active ingredients from highland barley have received considerable attention as natural products for developing treatments and dietary supplements against obesity.In practical application,the research of food combinations is more significant than a specific food component.This study investigated the lipid-lowering effect of highland barley polyphenols via lipase assay in vitro and HepG2 cells induced by oleic acid(OA).Five indexes,triglyceride(TG),total cholesterol(T-CHO),low density lipoprotein-cholesterol(LDL-C),aspartate aminotransferase(AST),and alanine aminotransferase(ALT),were used to evaluate the lipidlowering effect of highland barley extract.We also preliminary studied the lipid-lowering mechanism by Realtime fluorescent quantitative polymerase chain reaction(q PCR).The results indicated that highland barley extract contains many components with lipid-lowering effects,such as hyperoside and scoparone.In vitro,the lipase assay showed an 18.4%lipase inhibition rate when the additive contents of highland barley extract were 100μg/m L.The intracellular lipid-lowering effect of highland barley extract was examined using 0.25 mmol/L OA-induced HepG2 cells.The results showed that intracellular TG,LDL-C,and T-CHO content decreased by 34.4%,51.2%,and 18.4%,respectively.ALT and AST decreased by 51.6%and 20.7%compared with the untreated hyperlipidemic HepG2 cells.q PCR results showed that highland barley polyphenols could up-regulation the expression of lipid metabolism-related genes such as PPARγand Fabp4.

Metagenomic analysis revealing the metabolic role of microbial communities in the free amino acid biosynthesis of Monascus rice vinegar during fermentation

Free amino acid(FAA)is the important component of vinegar that infl uences quality perception and consumer acceptance.FAA is one of the major metabolites produced by microorganisms;however,the microbial metabolic network on FAA biosynthesis remains unclear.Through metagenomic analysis,this work aimed to elucidate the roles of microbes in FAA biosynthesis during Monascus rice vinegar fermentation.Taxonomic profiles from functional analyses showed 14 dominant genera with high contributions to the metabolism pathways.The metabolic network for FAA biosynthesis was then constructed,and the microbial distribution in different metabolic pathways was illuminated.The results revealed that 5 functional genera were closely involved in FAA biosynthesis.This study illuminated the metabolic roles of microorganisms in FAA biosynthesis and provided crucial insights into the functional attributes of microbiota in vinegar fermentation.

Lipids,lipid-lowering drug and sepsis:a Mendelian randomization study

lipid-lowering interventions on the disease.Methods:Two-sample Mendelian randomization analyses were conducted to evaluate the associations of high-density lipoprotein cholesterol,low-density lipoprotein cholesterol,triglycerides,apolipoprotein B and apolipoprotein A-I levels with risks for sepsis,and those of low-density lipoprotein cholesterol(HMGCR,PCSK9,NPC1L1),triglycerides(LPL,ANGPTL3,APOC3)and high-density lipoprotein cholesterol(CETP),apolipoprotein A-I(CETP),apolipoprotein B(HMGCR,PCSK9,NPC1L1,LPL,APOC3)with sepsis.Results:HMGCR-mediated low-density lipoprotein cholesterol and apolipoprotein B were associated with an increased risk of sepsis,with an odds ratio value of 1.4(95%confidence interval(CI):1.06-1.84,P=0.017)and 1.41(95%CI:1.01-1.98,P=0.046).CETP-mediated high-density lipoprotein cholesterol and apolipoprotein A-I were associated with a reduced risk of sepsis,with an odds ratio of 0.87(95%CI:0.82-0.92,P<0.01)respectively and 0.84(95%CI:0.78-0.9,P<0.01).Sensitivity analysis showed that the results were robust.Conclusion:HMG-CoA reductase inhibitors and CETP inhibitors may contribute to the prevention and treatment of sepsis.

N^+离子注入选育色素产生菌Monascus的研究

应用 1 0keV的N+离子注入,对产红色素红曲菌进行了诱变选育。在确定最佳注入剂量后,经过两轮离子注入诱变筛选,选育出 1株高产突变株,红色素色价提高 3 8.1 %,且色调适宜。经传代培养,其高产性状能稳定遗传。对该菌株的发酵特性也进行了研究。

氨基酸对于Monascus ruber生物合成Monacolin K影响的研究

在Monacolin K发酵中添加氨基酸后发现较高质量浓度的氨基酸高度抑制了Monacolin K的产量。0.1 g.L-1的D-甲硫氨酸在发酵第4 d添加可以提高产量30%以上,而L-甲硫氨酸则没有此功能,D,L-甲硫氨酸因D-甲硫氨酸的关系也有一定的增产效果。以甲硫氨酸代替蛋白胨作为主要氮源,则抑制了polyketide途径的前期步骤,因此严重抑制了色素及Monacolin K的生产。另外,发现1 g.L-1的L-苯丙氨酸的添加时间越早越有利于Monacolin K的生产,在起始时添加发酵单位可达135.9 mg.L-1,可能是因为L-苯丙氨酸经过脱氨后,可以进入polyketide途径从而促进了Monacolin K的生产。

红曲霉(Monascus anka As 3.782)菌种的紫外诱变和产色素的条件优化

应用红曲霉Monascus anka As3.782(CCTCC编号:AF93208)菌株,探索其固态发酵产红曲色素的最佳基质初始含水量,结果表明:含水量为约30%最好;通过正交试验优化其液态发酵产红曲色素的培养基,大米粉9%、NaNO33%、KH2PO40.3%、MgSO40.1%,黄豆粉0.5%,pH4.0时的培养基配方产红曲色价最高,达77.3U/mL;在紫外诱变距离20cm诱导3min的条件下筛选优良菌株,其固态发酵产红曲色价可达1156.667U/mL,是原始出发株360.667U/mL的3倍多,其液态发酵产红曲色价比原始出发株提高了2倍。

响应面法优化红曲霉菌株Monascus purpureusWX液态发酵产Monacolin K工艺条件

通过将单次单因子法和响应面法相结合的方法,对红曲霉菌株Monascus purpureusWX液态发酵产Monacolin K的工艺条件进行了优化.其较佳的工艺条件为:甘油102.2 g/L,黄豆粉29.2 g/L,NaNO32.0 g/L,MgSO4.7H2O 1.0 g/L,KH2PO41.0 g/L,初始pH 4.5,26℃下培养15 d.在此条件下,Monacolin K质量浓度达到297.4 mg/L,比优化前提高了约3.9倍.

红曲霉Monascus sanguineus X1处理黄水的培养基优化及酯化液制备

黄水是白酒酿造的主要副产物之一,富含多种有机质,可用于制备酯化液促进白酒增香,提高其利用率。研究了一株高产酯化酶的红曲霉菌株Monascus sanguineus X1,以酯化酶活力为指标,从碳源、碳氮比、接种量和pH值4个方面对X1菌株的培养基进行单因素实验及正交试验优化;采用酯化酶液处理黄水,制备酯化液,研究了黄水、乙醇、己酸等酯化前体物质添加量对酯化液制备的影响。结果表明,该红曲霉优化发酵培养基组分(以100 mL计):大米粉7.0 g,大豆蛋白胨2.0 g,NaNO30.2 g,KH2PO40.15 g,MgSO4·7H2O 0.1 g,培养基初始pH值5.0,接种量10%(体积分数)。在此条件下,添加体积分数为10%的黄水和体积分数为4%的乙醇,酶活力可达745.80 U/mL。向酯化酶液中加入体积分数为0.8%的己酸和体积分数为20%的乙醇继续培养1 d后,酯化液中己酸乙酯的体积分数可达0.121%;而向酯化酶液中补加体积分数为10%的黄水后,己酸乙酯和乳酸乙酯的体积分数分别可达0.055%和0.032%。该结果旨在为将黄水资源用于白酒增香提供理论参考。

一株产黄色素红曲霉Monascus HB-5的生物学特性及发酵条件研究(英文)

研究了一株产黄色素的红曲霉菌株Monascus HB-5的生物学特性,并对其接种量、pH、碳源、氮源等主要发酵因素进行优化。结果表明该菌产出高浓度单一黄色素,色调为棕黄色,仅在370nm具有吸收峰。产黄色素最适发酵条件为接种量10%,pH4.6～5.7,温度30～32℃,适合的碳源为玉米粉,氮源为硝酸铵,摇瓶黄色素色价达200 U/ml。并对色素的稳定性及安全性进行了初步研究,橘霉素含量低于0.1mg/l。

高产洛伐他汀红曲霉(Monascus sp.UV-D-9)的发酵条件优化

为了进一步提高洛伐他汀产量,对实验组前期获得的洛伐他汀高突变株Monascus sp.UV-D-9的发酵条件进行了响应曲面优化。获得最佳发酵条件为:发酵温度30.4℃,装液量134 m L,转速187 r·min-1,其他条件为发酵时间10 d,接种量8%。在此条件下,洛伐他汀的产量为(0.32±0.014)mg·m L-1(n=3),比原始条件的产量提高了10.1%。

Monascus ruber 5031洛伐他汀合成酶基因的PCR分析

目的分析Monascus ruber5031洛伐他汀合成酶基因的结构。方法用3对以土曲霉洛伐他汀合成酶基因为基础设计的PCR引物对红曲霉基因组进行了PCR分析,其中一对引物序列为,正向:5’TTC GAT GCG GCC TTC TTC AAT3’和反向5’ATG GTT CGG CATCGT AAT TCC 3’。结果在红曲霉基因组中相当于土曲霉洛伐他汀合成酶基因的LNKS区域扩增出了745 bp的核酸片段,其序列与土曲霉合成酶的这一区域序列相同。结论土曲霉和红曲霉洛伐他汀合成酶基因存在同源区域。

产红色素真菌Monascus sanguineus的分离及其色素提取条件研究

采用组织块分离法首次从药用植物地黄中分离出一株产红色素内生真菌RJL03,根据形态学特征和r DNA ITS1-5.8S-ITS2序列分析的手段鉴定为Monascus sanguineus。采用响应面法(RSM)对菌株RJL03中红色素提取工艺条件进行研究。在单因素实验的基础上,以提取液OD值为指标,利用Box-Behnken中心组合实验设计原理(BBD)优化菌株中红色素的有机溶剂超声波辅助浸提(UAE)工艺。结果显示,该色素提取液在515 nm处有最大吸收波长,最佳提取工艺为温度70℃、提取时间34 min、乙醇浓度50%。在此条件下提取菌株中的红色素,得到的提取液OD值为2.587,可比优化前提高约1.5倍。

产红色素真菌Monascus sanguineus的液态发酵条件研究

从药用植物地黄中分离出一株产红色素内生真菌RJL03(Monascus sanguineus),为进一步提高菌株RJL03的红色素产量,采用响应面法(RSM)对其液态摇瓶发酵条件进行研究。在单因素实验的基础上,以发酵液OD值为指标,利用Box-Behnken中心组合实验设计原理(BBD)优化菌株液态发酵产红色素的工艺条件。结果显示,该色素发酵液在474 nm处有最大吸收波长,最佳发酵工艺为温度30℃、装瓶量67%、初始p H自然(7)。在此条件下液态发酵菌株,显著提高了菌株RJL03的红色素产量。

高产γ-氨基丁酸低产桔霉素红曲霉(Monascus ruber)突变子1047筛选与发酵特性研究

以红曲米中筛选到的红色红曲霉菌株G为原始菌株,通过农杆菌介导T-DNA插入突变技术,成功构建了含有1 483株红曲霉突变子的T-DNA插入突变库。用HPLC等方法从突变库中筛选出10株γ-氨基丁酸(GABA)产量稳定高于原始菌株G的突变子,薄层层析结合HPLC技术分析了10株突变子发酵液中桔霉素的含量;其中突变子1047的GABA产量较高,为1.169g/L,是原始菌株G(GABA产量0.472g/L)的247.67%,桔霉素含量稳定低于原始菌株G。以红曲霉菌株G和突变子1047为实验材料,通过5 L发酵罐发酵并定时取样,HPLC等方法精确分析发酵液各种活性物质的含量;结果显示,突变子1047生长速度稍快于原始菌株G;GABA、莫纳可林K(Monacolin K)、红曲色素色价分别为2.201 g/L、83.892 mg/L、21.984 U/mL,是原始菌株G的279.67%、108.01%和182.35%;而桔霉素产量为1.976 mg/L,是原始菌株G的41.71%。因此,利用TDNA插入的方法对红曲霉进行育种,能产生稳定的遗传变化,在红曲霉资源的保护和利用上有一定潜力。

怀地黄内生产红色素真菌血红红曲霉(Monascus sanguineus)的分离与鉴定

真菌色素是天然食用色素的潜在来源。本文从药用植物怀地黄(Rehmannia glutinosa Libosch.)中分离鉴定出了一株能够产生丰富可溶性红色素的内生红曲霉属真菌RJL03。为进一步研究和利用该菌株及其次级代谢物,对其进行分类鉴定。采用不同的培养基培养并观察RJL03的菌落特征,使用光学显微镜和扫描电子显微镜对菌株形态进行观察发现:该菌株菌落初为白色,后产色素变红,绒毡状,圆形;菌丝分枝,分隔;分生孢子倒梨形,单生或至多10个成短链状,大小为(8.0～16.5)μm×(7.5～14.0)μm;孢子囊球形,外壳褐色,直径32～70μm;子囊孢子椭圆形,大小为(6.0～7.5)μm×(4.0～5.0)μm,表面平滑,无色或红色。序列分析发现,该菌株ITS和18S rDNA序列与血红红曲霉(Monascus sanguineus)的相似度分别为100%和93%。综合形态学和序列分析的结果,将菌株RJL03鉴定为血红红曲霉。这是首次在我国报道的血红红曲霉怀地黄分离株;另外,通过对比观察认为M.sanguineus是与紫色红曲霉(M. purpureus)不同的独立物种。

pH对红色红曲菌(Monascus ruber)M7生长及产孢的影响

研究红色红曲菌(Monascus ruber)M7生长、繁殖两个不同阶段对环境pH需求的差异,在孢子培养过程中对pH进行分段控制,旨在减小产孢条件对菌丝生长的影响,提高孢子产量。结果表明,菌株M7孢子在pH 3.0~6.0条件下培养8 h均可100%萌发,其中,pH4.0~5.0最佳,萌发最为迅速,在pH 7.0~8.0时孢子萌发受到严重抑制,萌发率很低;菌落生长在pH 4.0~6.0时较快,其中,pH 5.0最佳,菌落直径最大,但在pH 3.0和7.0时菌落生长明显减慢,在pH 8.0时受到严重抑制,形成的菌落很小;pH>8.0的碱性条件为极端pH条件,孢子萌发和菌落生长完全被抑制。在菌株M7生长的pH范围内,产孢能力随着pH的升高而提高,适宜的产孢pH范围为7.0~8.0。根据菌株M7生长和繁殖对pH的需求差异,控制培养前期pH值为5.0、后期pH值为7.0~8.0,在PDB和CYB培养基中的产孢量分别提高了12~16倍和3~4倍。

Cloning and Sequence Analysis of the Full-length cDNA of a Novel yp05 Gene Associated With Citrinin Production in Monascus aurantiacus

Objective To obtain the full-length cDNA of a novel gene （named yp05） associated with citrinin production-related genes in Monascus aurantiacus. Methods Total RNA was extracted from mycelium, 3＇ and 5＇ cDNA end of yp05 gene was amplified using smartTM trace cDNA amplification kit, and the full-length cDNA of a novel gene （named yp05） was obtained from the electronic assembly of 3＇-RACE and 5＇- RACE products. Results This yp05 gene was 787 bp including a 597 bp open reading frame （ORF） and encoded a deduced protein with 199 amino acid residues, and the amino acid sequence of this protein was found similar with the sequences of many fungal manganese-superoxide dismutases in the GenBank with the aid of BLASTp. The transcription ofyp05 gene in Monascus strains was analyzed with the aid of Northern blotting. The transcription of yp05 gene was only detected in Monascus strains, provided that citrinin was produced. Conclusion The transcription of yp05 gene belongs to differential expression genes of citfinin yielded from Monascus and has no correlation with the biosynthesis pathway of red pigments.

Serial Analysis of Gene Expression in Monascus aurantiacus Producing Citrinin

To construct a tag expression library of Monascus aurantiacus that could produce citrinin maximally on the thirteenth (0.966 mg/mL) day in the submerged culture. Methods Total RNA was extracted from the mycelium, cDNA was synthesized using the SuperScript choice system, and then, a SAGE library was successfully constructed according to the MicroSAGE method. Results Five hundred and ninety eight clones were obtained in SAGE library, and 120 clones were picked out randomly for identification and sequencing purpose. Eighty nine clones had positive inserts, 26 clones had no inserts and the remaining 5 clones had no site of NlaⅢ enzyme in inserts. There were seven repeated clones. Conclusion With the aid of SAGE2000 software, 901 tags were obtained from 89 clones, representing 686 unique transcripts. Six unique tags of them belong to highly expressed genes (Number of tags≥10) and 143 unique tags to moderately expressed genes (repeat tags≥2).

Monascus pilosus-fermented black soybean inhibits lipid accumulation in adipocytes and in high-fat diet-induced obese mice

Objective:To explore the anti-obesity effects and the mechanism of action of Monascus pilosus(M.pilosus)-fermented black soybean(MFBS)extracts(MFBSE)and MFBS powders(MFBSP)in adipocytes and high-fat diet(HFD)-induced obese mice,respectively.Methods:Black soybean was fermented with M.pilosus,and the main constituents in MFBS were analyzed by HPLC analysis.In vitro,MFBSE were examined for anti-adipogenic effects using Oil-Red O staining.In vivo,mice were fed a normal-fat diet(NFD)control,HFD control or HFD containing 1 g/kg MFBSP for 12 weeks,and then body weight gain and tissues weight measured.Real-time PCR and western blot assay were used to determine the mechanism of anti-adipogenic effects.Results:MFBSE inhibited lipid accumulation in 3T3-L1 adipocytes without exerting cell cytotoxicity.MFBSP treatment in HFD-fed mice significantly decreased the body weight gain compared with the HFD control mice.MFBSE and MFBSP treatment resulted in significantly lower mRNA levels of adipogenesis-related genes,such as peroxisome proliferator-activated receptorγ(PPARγ),fatty acid-binding protein 4(FABP4),and fatty acid synthase(FAS),in adipocytes and in white adipose tissue(WAT)of HFD-induced obese mice.Conclusions:These results suggest that the anti-obesity effects of MFBS are elicited by regulating the expression of adipogenesis-related genes in adipocytes and WAT of HFDinduced obese mice.

Extracts of Monascusus Purpureus beyond Statins——Profile of Efficacy and Safety of the Use of Extracts of Monascus Purpureus

Extracts of Monascus purpureus have always been considered a natural source of Iovastatin, the precursor of the world＇s largest selling class of drugs. In actual fact, the fungus contains many other substances （flavonoids, polyunsaturated fats, pyrrolinic compounds etc.） with a wide variety of other actions. The most recent studies have shown that it has an action on the glycemic metabolism, and on the mechanisms of adipogenesis, also an effects on the endothelium and on postprandial vasodilation. These effects are more extensive and complex than those of statins alone. And new strains of Monascus purpureus have recently been patented where the presence of statins is only one of the therapeutic components of the fungus. In particular, the increase in secondary components, such as flavonoids, which coincides with a more complex therapeutic action, probably making the new extracts of Monascus purpureus, the ideal candidate for the treatment of the metabolic syndrome.