新型固定化Aspergillus oryzae脂肪酶催化合成1,3-二油酸-2-棕榈酸甘油三酯

旨在评价自制固定化sn-1,3专一性脂肪酶Aspergillus oryzae L03(AOL)替代商业固定化脂肪酶酸解法合成1,3-二油酸-2-棕榈酸甘油三酯(1,3-dioleoyl-2-palmitoylglycerol,OPO)的可行性。以棕榈硬脂和油酸为原料,在正己烷体系中,得到AOL最适反应参数为:反应温度55℃,反应时间1 h,棕榈硬脂和油酸的底物摩尔比为1∶8,正己烷与底物混合物质量比1∶6,AOL水分含量3.5%(w/w)。该条件下,OPO含量44.9%,三棕榈酸甘油酯(PPP)含量为3.23%,sn-2位棕榈酸占所有棕榈酸比例为68.86%,符合国家标准。同时,AOL和LipozymeRM IM、LipozymeTL IM两种商品酶对比,AOL的催化速率最快,1 h达到最大OPO生产峰值。经过8次循环使用,AOL具有良好的操作稳定性。研究表明,自制的新型廉价固定化酶AOL具有良好的应用前景。

Nucleotide oligomerization domain 2 contributes to the innate immune response in THCE cells stimulated by Aspergillus fumigatus conidia

AIM: To investigate the expression of nucleotide oligomerization domain 2 (NOD2) in the immortalized human corneal epithelial cell line (THCE), and its role in the innate immune response triggered by inactive Aspergillus fumigatus (Af) conidia. METHODS: The normal THCE cells were investigated as controls. After incubation with inactive Af conidia for 0.5, 2, 4, 6, and 8 hours, THCE cells were harvested, mRNA expression of NOD2 and receptor interacting protein 2 (RIP2) was detected by RT-PCR. Intracellular proteins including NOD2, NF-kappa B and proinflammatory cytokines such as TNF-alpha, IL-8, IL-6 in the cell supernatant were analyzed by ELISA. RESULTS: Our data indicate that NOD2 expressed in the normal THCE cells. After triggered by the inactive Af conidia, the expression of NOD2, RIP2 mRNA and the secretion of NOD2, NF-kappa B, TNF-alpha, IL-8, IL-6 both increased in a time-depended manner, and reached the peak point at 4, 6, 6, 4, 6, 6, 4 hours, respectively. And after pretreated with NOD2 neutralizing antibody, the expression of RIP2, NF-kappa B, TNF-alpha, IL-8 both decreased dramatically at the peak point, while the secretion of IL-6 changed little. CONCLUSION: The results of this study suggest that NOD2 exists and expresses in the THCE cells, and contributes to the innate immune responses triggered by inactive Afconidia by induction of proinflammatory cytokines such as TNF-alpha and IL-8 through the NF-kappa B pathway.

Role of the IL-33/ST2/p38 signaling pathway in the immune response of corneal epithelial cells to Aspergillus fumigatus infection

AIM: To investigate the expression of interleukin(IL)-33 in the cornea and human corneal epithelial cells(HCECs) exposed to Aspergillus fumigatus(A. fumigatus), and to determine the function of IL-33/ST2/p38 signaling pathway in the immune response of corneal epithelial cells to A. fumigatus infection.METHODS: The mRNA and protein expression of IL-33 in HCECs and mice corneas were examined by quantitative real-time reverse transcription polymerase chain reaction(q RT-PCR) and Western blot analysis, respectively. IL-33 expression was also detected in cornea samples from healthy donors and patients with fungal keratitis with immunohistochemistry. The cultured HCECs were treated with inactive A. fumigatus hyphae at various concentrations with or without recombinant human IL-33 protein, soluble recombinant ST2 protein, specific ST2 neutralizing antibody, or the mitogen-activated protein kinase(MAPK) p38 inhibitor SB203580 for evaluation of the expression and activation of IL-33/ST2/p38 signaling in the regulation of proinflammatory cytokines. The production levels of IL-6 and IL-1β were determined by qR T-PCR and enzymelinked immunosorbent assay(ELISA). The proliferation of HCECs was determined by a Cell Counting Kit-8(CCK8) assay and cell count.RESULTS: IL-33 expression levels increased in the corneal tissues of patients with fungal keratitis and in mice corneas of experimental A. fumigatus infection,as well as in HCECs with infection of A. fumigatus. A. fumigatus strongly stimulated HCECs-generated proinflammatory cytokine(IL-6 and IL-1β) production at both the mRNA and protein levels. This production of proinflammatory mediators stimulated by A. fumigatus was further stimulated by IL-33 and was prevented by soluble ST2 protein or ST2 neutralizing antibody. Moreover, IL-33 naturally promoted the p38 phosphorylation induced by A. fumigatus, which was suppressed by soluble ST2 protein. The MAPK p38 inhibitor SB203580 also inhibited the A. fumigatus-induced proinflammatory cytokine production. IL-33 administration for 48 h and 72 h promoted the proliferation of HCECs, which was attenuated by treatment with soluble recombinant human ST2 protein.CONCLUSION: A. fumigatus elevates IL-33 expression in human and mice corneas and HCECs. Thus, IL-33/ST2/p38 signaling may play an important role in amplifying the immune response of corneal epithelial cells to A. fumigatus infection. Besides, IL-33 promotes the cell proliferation of HCECs via its receptor ST2. These findings suggest a novel autocrine mechanism of amplification of the fungalinduced inflammatory response in the corneal epithelium, highlighting a potential therapeutic target for fungal keratitis.

Pumpkin-like MoP-MoS_(2)@Aspergillus niger spore-derived N-doped carbon heterostructure for enhanced potassium storage

Biomass-derived carbon materials are widely applied in the energy storage and conversion fields due to their rich sources,low price and environmental friendliness.Herein,a unique pumpkin-like MoPMoS_(2)@Aspergillus niger spore-derived N-doped carbon(SNC)composite has been prepared via a simple hydrothermal and subsequent phosphorization process.Interestingly,the resulting MoP-MoS_(2)@SNC well inherits the pristine morphology of spore carbon,similar to the natural pumpkin,with hollow interiors and uneven protrusions on the surface.The special structure allows it to have sufficient space to fully contact the electrolyte and greatly reduces the ion transport distance.The theory calculations further demonstrate that the formed MoP-MoS_(2)heterostructure can enhance the adsorption of K ions and electronic couplings.With these unique advantages,the MoP-MoS_(2)@SNC anode for potassium storage shows a high reversible capability of 286.2 mAh g&(-1) at 100 mA g^(-1) after 100 cycles and superior rate performance.The enhanced electrochemical performance is mainly related to the unique pumpkin-like morphology of SNC and the construction of MoP-MoS_(2)heterostructure,as well as their perfect coupling.This study provides a feasible design idea for developing green,low-cost,and high-performance electrode materials for next-generation energy storage.

Expression of indoleamine 2,3-dioxygenase in a murine model of Aspergillus fumigatus keratitis

AIM： To observe the presence and expression of indoleamine 2,3-dioxygenase（IDO） during the corneal immunity to Aspergillus fumigatus（A. fumigatus） in the murine models.·METHODS： The murine model of fungal keratitis was established by smearing with colonies of A. fumigatus after scraping central epithelium of cornea and covering with contact lenses in C57BL/6 mice. The mice were randomly divided into control group, sham group and A.fumigatus keratitis group. The cornea was monitored daily using a slit lamp and recorded disease score after infection. Corneal lesion was detected by immunofluorescence staining. IDO m RNA and protein were also detected by quantitative reverse transcription-polymerase chain reaction（q RT-PCR） and Western blot.· RESULTS： The disease score and slit lamp photography indicated that disease severity was consistent with corneal inflammation in the murine models, and the disease scores in A. fumigatus keratitis group were obviously higher than those in the sham group. By immunofluorescence staining, IDO was mainly localized in corneal epithelium and stroma in the murine corneal tissues with A. fumigatus keratitis. Compared with the sham group, IDO m RNA expression was significantly enhanced in corneal epithelium infected by A. fumigatus. Furthermore, IDO protein expression detected by Western blot was in accord with transcript levels of IDO m RNA measured by q RT-PCR. IDO protein expression was enhanced after A. fumigatus infection compared with the sham group.·CONCLUSION： IDO is detected in corneal epithelium and stroma locally, which indicates IDO takes part in the pathogenesis of A. fumigatus keratitis and plays a key role in immune regulation at the early stage.

Indoleamine 2,3-dioxygenase adjusts neutrophils recruitment and chemotaxis in Aspergillus fumigatus keratitis

AIM: To explore the effect of indoleamine 2,3-dioxygenase(IDO) on recruitment and chemotaxis function of neutrophils in Aspergillus fumigatus(A.fumigatus) keratitis.METHODS: C57BL/6 mice models of A.fumigatus keratitis were established by inoculating hyphae of A.fumigatus evenly on the corneas.The clinical scores and inflammatory cytokines expression were measured respectively on the 1^(st), 3^(th), 5^(th) day after infection.The 1-MT(1 mg/m L) was administered by gavage to exert an inhibitory effect on IDO during infection.The mice were divided into control group, 1-MT group, A.fumigatus(A.F.) group, and 1-MT+A.F.groups.The corneas were monitored by slit lamp microscopy, and recorded disease scores in 3 d after infection.Myeloperoxidase(MPO) assay was done to evaluate the neutrophils infiltration.Immunofluorescence staining was used to detect the recruitment of neutrophils in murine corneas.The m RNA of inflammatory cytokines was measured with reverse transcription-polymerase chain reaction(RT-PCR).RESULTS: The corneal inflammation and the clinical score reached the peak on the 3;day after the corneal infection.The m RNA of inflammatory cytokines of the A.F.group reached the highest on the 3;day after the infection accordingly.Meanwhile, the results of slit light photography indicated that inhibitors of IDO made inflammation more serious contrasted with the A.F.group on the 3;day.Besides, imunofluorescence staining and MPO indicated that 1-MT enhanced the recruitment, infiltration and chemotaxis of neutrophils obviously in contrast to the A.F.group.RT-PCR indicated that 1-MT increased the expression of CXCL-1, ICAM-1, IL-1β, and IL-8 significantly.CONCLUSION: IDO participates in the pathogenesis of A.fumigatus keratitis and plays an important role in inducing immune protection by inhibiting neutrophils-related inflammatory reaction and suppressing recruitment and chemotaxis of the neutrophils.

真菌Aspergillus fumigatus var .fumigatus的次生代谢产物(英文)

目的:研究西双版纳粗榧共生真菌Aspergillus fumigatusvar .fumigatus的次生代谢产物。方法:通过柱层析法分离发酵产物的化学成分,并利用波谱数据和文献数据对照鉴定化合物的结构。结果:分离到2个化合物,分别鉴定为trypacidin (1)及其衍生物1 ,2-seco-trypacidin (2)。结论:化合物2是新结构;并对文献中关于trypacidin的C-4和C-7a位碳谱数据的指定进行了更正。

黑曲霉Aspergillus niger对人参皂苷Re的微生物转化

目的:筛选长白山人参土壤中的活性微生物,转化单体人参皂苷产生稀有人参皂苷成份。方法:从长白山人参根际土壤中分离各类菌株,对单体人参皂苷Re进行微生物转化,通过硅胶柱层析等方法对转化产物进行分离纯化,采用波谱解析及理化常数对其进行结构鉴定。结果:从长白山人参根际土壤中分离各类真菌菌株68株,3株真菌对三醇组人参皂苷Re具有转化作用,其中黑曲霉Aspergillusniger的转化活性较强,转化产物为人参皂苷Rg1、Rg2和Rh1。结论:首次报道黑曲霉能将人参皂苷Re转化为人参皂苷Rg1、Rg2和Rh1这一转化过程。

蛋白组学差异揭示黑曲霉(Aspergillus niger)高产柠檬酸的分子机制

黑曲霉(Aspergillus niger)YX-1217是一株典型的柠檬酸高产菌株,基于二维聚丙烯酰胺凝胶电泳(2-DE)对黑曲霉YX-1217和退化菌株YX-1217G的胞外分泌蛋白进行比较,同时利用同位素标记相对和绝对定量(i TRAQ)技术比较了两株菌的胞内蛋白差异,揭示黑曲霉YX-1217高产柠檬酸的分子机制。结果显示,菌株黑曲霉YX-1217G的胞外蛋白数量和浓度均显著低于菌株黑曲霉YX-1217(P<0.05),通过质谱鉴定了6个黑曲霉YX-1217高表达蛋白;利用i TRAQ技术共鉴定到3 553个蛋白,将差异蛋白按照京都基因与基因组百科全书(KEGG)代谢数据库进行分类,发现与柠檬酸生物合成相关的酶在菌株黑曲霉YX-1217中的表达量显著高于退化菌株(P<0.05),这些蛋白的高效表达是菌株黑曲霉YX-1217高产柠檬酸的原因之一,为开发柠檬酸或其他有机酸的高产菌株提供新的思路和观点。

Metabolites of Marine Fungus Aspergillus sp. Collected from Soft Coral Sarcophyton tortuosum

Fungus Aspergillus sp., which was isolated from soft coral Sarcophyton tortuosum, was cultured in the GPY medium containing glucose 10 g/L, peptone 5 g/L, yeast extract 2 g/L, sea water 1 L, at pH=7.5. Four com pounds, 3,6-diisobutyl-2（1H）-pyrazinone（1）, 3-isobutyl-6-（1-hydroxy-2-methylpropyl）-2（1H）-pyrazinone（2）, 3-methoxy-4-methyl-2,4-dien-pentanoic acid（3） and penicillic acid（4） were obtained from the AcOEt extract of the culture broth. Their structures were elucidated mainly based on the NMR, HR-EI-MS and X-ray single crystal diffraction experimental data. Compound 3 is a new compound. Compound 1 was previously proposed to be the tautomer of flavacol（3,6-diisobutylpyrazin-2-ol, 5）. However, the evaluation of the NMR and X-ray single crystal diffraction experimental data permitted us to propose that compound 1 existed as amide form instead oftautomers. Compound 1 is stable both in solution and crystal due to strong intermolecular hydrogen bonds. Incidentally supplying anthracenone to the GPY medium with a dose of 100 mg/L, the yield of penicillic acid（4） in the culture broth of the strain Aspergillus sp. was increased dramatically from 6 mg/L to 57 mg/L.

A new naphthoquinoneimine derivative from the marine algal-derived endophytic fungus Aspergillus niger EN-13

Cultivation of an endophytic fungus Aspergillus niger EN-13 that was isolated from the inner tissue of the marine brown alga Colpomenia sinuosa resulted in the characterization of a new naphthoquinoneimine derivative, namely, 5,7-dihydroxy-2-[1-（4- methoxy-6-oxo-6H-pyran-2-yl）-2-phenylethylamino]-[ 1,4]naphthoquinone. The structure of the new compound was established on the basis of various NMR spectroscopic analyses including 2D NMR techniques, El-MS, and HR-ESI-MS. This compound displayed moderate antifungal activity.

Vanillin and its derivatives,potential promising antifungal agents,inhibit Aspergillus flavus spores via destroying the integrity of cell membrane rather than cell wall

To study the structure-antifungal activity of vanillin against Aspergillus flavus(A.flavus),the susceptibilities of A.flavus to vanillin and its isomers(o-vanillin,2-hydroxy-4-methoxybenzaldehyde(HMB),2-hydroxy-5-methoxybenzaldehyde)and the possible antifungal mechanisms have been investigated.All the four volatile aldehydes could inhibit the germination of spores,and the minimum inhibitory concentrations(MICs)of them were in this order:vanillin(200μg/mL),o-vanillin(100μg/mL),2-hydroxy-5-methoxybenzaldehyde(100μg/mL),HMB(70μg/mL).The minimum fungicidal concentrations(MFCs)of them were in this order:vanillin(240μg/mL),o-vanillin(160μg/mL),HMB(140μg/mL),2-hydroxy-5-methoxybenzaldehyde(140μg/mL).Spore size was arrested at 0 h with the treatment of the four volatile aldehydes.Effects of the four volatile aldehydes on the cell wall and cell membrane integrity of A flavus were observed by calcofluor white(CW)staining and propidium iodide(PI)staining.The results showed that HMB exerted the strongest antifungal and fungicidal effects on the growth of A.flavus.The four volatile aldehydes had little influence on cell wall integrity after 3-hour treatment,however,they could strongly damage the cell membrane integrity.All the four volatile aldehydes could effectively prevent the growth of A.flavus on peanut seeds.The antifungal mechanisms of the four volatile aldehydes provide theoretical foundations for their development of new antifungal agents.

枯草芽孢杆菌PW2挥发性产物对黄曲霉的抑制作用

为探寻安全、高效的黄曲霉新型抑制物,采用顶空固相微萃取-气质联用(HS-SPME-GC-MS)技术对枯草芽孢杆菌(Bacillus subtilis)PW2所产具有抗霉活性的挥发性有机化合物(volatile organic compounds,VOCs)进行了成分鉴定,并利用平板对扣法从鉴定出的成分中筛选具有强抗霉活性的目标化合物,通过平板对扣法和96孔板梯度稀释法测定目标化合物对黄曲霉的最小抑菌体积分数(MIC),使用黄曲霉毒素B_(1)(aflatoxin B_(1),AFB_(1))ELISA检测试剂盒分析了目标化合物对黄曲霉产AFB_(1)的抑制作用,使用扫描电子显微镜(scanning electron microscope,SEM)观察受试黄曲霉孢子形态的变化和采用分光光度法测定受试黄曲霉细胞膜麦角甾醇相对含量,以探讨抗霉机理。结果显示,从PW2所产VOCs中共鉴定出41种组分,并从中筛选出异辛醇为目标化合物;平板对扣法和96孔板梯度稀释法测得异辛醇对黄曲霉的MIC分别为0.169μL/mL和6.25μL/mL;PDB培养基中异辛醇体积分数分别为3.13、6.25、12.50μL/mL时,均能有效抑制AFB_(1)的产生,且随异辛醇体积分数的升高,抑制效果增强;异辛醇处理可导致黄曲霉孢子表面凹陷和孢子破裂,并且影响细胞膜中麦角甾醇的相对含量,造成细胞膜受损。该研究可为开发以异辛醇为功能成分的黄曲霉抑制剂提供理论参考。

The Difference in Calcium Levels in Aspergillus nidulans Grown on Glucose or Pectin

Understanding the growth regulatory mechanisms in filamentous fungi is very important for the production of medicines for antifungal therapies. It is well established that Ca2+ gradient is essential for hyphal growth and that one mechanism responsible for the Ca2+ cellular concentration starts with the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) by receptor-regulated forms of phosphoinositide-specific phospholipase C (PI-PLC). In the present study the levels of calcium in Aspergillus nidulans wild type (A26) and plcA-deficient mutant (AP27) growing in a carbon source readily assimilated, as glucose or pectin a non-readily assimilated carbon source was investigated. Intracellular calcium levels in A26 were higher in the presence of glucose than in pectin, but lower in AP27 independently of the carbon source and in AP27 the vesicular calcium distribution occurred mainly at the apex of the hyphae. Delay in nuclear division was also observed if A26 and AP27 were grown in pectin presence when compared with growth in glucose. For the first time, it is demonstrated that the levels of intracellular Ca2+ were higher when A. nidulans was growing in glucose than in a non readily assimilated carbon source as pectin. Further, it also showed that the plcA gene, although not essential, may be responsible for high-molecular weight carbon source recongnation, for the intracellular Ca2+ levels maintenance and consequently by the nuclear division in A. nidulans.

Interactions of thymic stromal lymphopoietin with interleukin-4 in adaptive immunity during Aspergillus fumigatus keratitis

AIM:To investigate the potential interactions of thymic stromal lymphopoietin(TSLP)with interleukin-4(IL-4)in adaptive immunity during fungal keratitis(FK).METHODS:An FK mouse model was induced with Aspergillus fumigatus(AF)hyphal infection.Mice were divided into several groups:untreated,phosphate buffer saline(PBS),infected with AF,and pretreated with a scrambled siRNA,a TSLP-specific siRNA(TSLP siRNA),murine recombinant TSLP(rTSLP),immunoglobulin G(IgG),murine recombinant IFN(rIFN-γ),murine recombinant IL-4(rI L-4),rIL-13,murine recombinant IL-17A(rIL-17A),and murine recombinant IL-17F(rIL-17F)groups.Quantitative realtime reverse transcription-polymerase chain reaction(qRTPCR)and enzyme-linked immunosorbent assay(ELISA)or Western blot were performed to determine mRNA and protein levels in the inflamed cornea.Cytokine locations were observed by immunofluoresence staining after AF hyphal infection.RESULTS:Compared to those in the untreated group,TSLP and T helper type 1(Th1)cytokine levels in the AF group were upregulated at 24 h post infection(hpi),and those of T helper type 2(Th2)and T helper type 17(Th17)cytokines were increased at 5 d post infection(dpi).Th2 cytokine levels were decreased in the TSLP siRNA-pretreated group and increased in the rTSLP-pretreated group compared with the AF group.The TSLP level was increased in the rIL-4-pretreated group,but there were no significant changes among the other groups.Immunofluorescence staining showed cytokine locations after AF hyphal infection.CONCLUSION:TSLP induces a Th2 immune response and promots Th2 T cell differentiation in vivo.IL-4 promotes TSLP secretion.Therefore,TSLP with IL-4 regulates adaptive immunity in FK.

冠突曲霉UBC与MAT1-2-1互作关系验证及功能研究

【目的】验证冠突曲霉泛素结合酶(UBC)与交配型蛋白(MAT1-2-1)的互作关系,对UBC基因进行功能研究,为深入解析冠突曲霉有性产孢机制提供理论参考。【方法】采用实时荧光定量PCR检测UBC基因在冠突曲霉有性发育阶段的表达水平,酵母双杂交试验验证MAT1-2-1与UBC的互作关系,利用无缝克隆方法构建pDHt/sk-hyg-UBC过表达载体,并转化冠突曲霉获得UBC过表达菌株,对过表达菌株进行表达量检测、形态学观察及氧和盐胁迫的耐受性测定。【结果】PCR克隆获得UBC基因的编码区(CDS)序列,长度为474 bp,编码157个氨基酸残基,相对分子质量为39587.24 Da,理论等电点为5.16,为疏水的不稳定蛋白,含跨膜结构域,定位于细胞核,属于UBCc超家族成员。有性发育阶段(闭囊壳形成期和子囊孢子大量形成期)UBC基因的相对表达量显著高于营养菌丝体阶段(P<0.05,下同)。自激活检测结果显示,UBC无自激活作用;酵母双杂交验证结果显示,UBC与MAT1-2-1存在互作关系。通过构建过表达载体pDHt/sk-hyg-UBC成功获得UBC过表达菌株(OE::UBC)。与野生型菌株相比,UBC和MAT1-2-1基因在UBC过表达菌株的相对表达量均显著升高。UBC过表达菌株与野生型菌株在MYA、MYA+5%NaCl和MYA+17%NaCl固体培养基上的菌落形态无明显差异。过表达菌株能分别在含18 mmol/L过氧化氢和25%NaCl的培养基中生长,而野生型菌株不能生长,说明UBC基因的过表达提高冠突曲霉对氧胁迫和盐胁迫的耐受性。【结论】UBC基因可能与冠突曲霉的有性发育相关,且与MAT1-2-1基因存在共表达特征。由于UBC与MAT1-2-1存在互作关系,且UBC基因的过表达可明显增强冠突曲霉的抗氧化和抗盐能力,故推测UBC通过与MAT1-2-1发生互作共同参与冠突曲霉的氧胁迫和盐胁迫应答过程。

Antifungal Activity against <i>Aspergillus parasiticus</i>of Supernatants from Whey Permeates Fermented with Kefir Grains

Aspergillus parasiticus, a common fungal contaminant in food, produces aflatoxin B1, which is classified as human carcinogen. Kefir is an ancient fermented beverage obtained by the fermentation of different substrates with kefir grains. A very important waste produced by the dairy cheese industry is the whey permeate, which nowadays is a strong ambient contaminant. The aim of this work was to assess the effect of whey permeates fermented with kefir grains against A. parasiticus growth, aflatoxin B1 biosynthesis, and the kefir microorganisms protection against the cell damage produced by aflatoxin B1. It was observed that kefir-cell-free-supernatants (CFS) produced fungal inhibition. A fungicidal effect was observed with 65% v/v of CFS in the culture medium (final pH 4.55 and total undissociated lactic and acetic acid concentration 34.08 mM). Under these conditions, aflatoxin production was not detected. Finally, it was found that non-viable kefir microorganisms protected HepG2 cells from the damage produced by aflatoxin B1.

核苷酸结合寡聚化结构域2在大鼠烟曲霉菌性角膜炎角膜局部的表达

目的探讨大鼠角膜真菌感染后角膜局部核苷酸结合寡聚化结构域2(NOD2)的表达。方法在大鼠左眼角膜上制作烟曲霉菌性角膜炎的模型(实验组),右眼作为对照组。在感染后第2天(早期)、第6天(中期)及第14天(晚期)取下角膜组织,采用逆转录聚合酶链反应(RT-PCR)检测NOD2在不同病程角膜组织中的表达。结果在实验组角膜中,NOD2早期开始表达,中期达高峰,以后逐渐下降。实验组与对照组早、中期NOD2 mRNA表达的差异均有统计学意义(P值均<0.05),晚期的差异无统计学意义(P>0.05);且实验组早期与中期、中期与晚期表达的差异均有统计学意义(P值均<0.01)。结论NOD2参与角膜烟曲霉菌性感染后角膜局部的免疫反应。

青霉T24-2和灰绿曲霉EU7-22降解甘蔗渣产糖的研究

青霉T24-2(Penicilliumsp.T24-2)和灰绿曲霉EU7-22(Aspergillus glaucusEU7-22)能利用甘蔗渣进行固态发酵产纤维素酶.通过对青霉和灰绿曲霉混合培养特性的研究,确定两个菌株不适合进行混合培养产纤维素酶.分别对两个菌株产纤维素酶的条件进行优化,单独用青霉曲酶时糖化率为31.4%,单独用灰绿曲霉曲酶时糖化率为34.1%.对其曲酶混合糖化进行研究,当两种曲酶以1∶1比例进行混合糖化时,糖化率可达51.3%.以青霉T24-2和灰绿曲霉EU7-22的曲霉混合糖化的研究国内外还未见报道,研究结果大大提高了糖化率.

无花果曲霉对Cu^(2+)吸附的研究

采用生物吸附法 ,研究了经过预处理的无花果曲霉对Cu2 + 吸附行为与影响吸附的主要因素 ,具体包括菌丝球与Cu2 + 接触反应时间、Cu2 + 浓度、原初pH值 3个方面。结果表明 :该菌丝球在吸附进行 5min时 ,吸附量已达到 2 .85 5mg/g ,在吸附进行 4h以后趋于平衡。最佳pH范围为 4.0 - 5 .0 ,Cu2 + 浓度在 2 0 - 10 0mg/L范围内吸附量为 2 .76 3 - 4.713mg/g ,而吸附率却为 32 .31% - 9.5 4%。