Objective:To study the effects of liquid culture conditions on inhibition of Paecilomyces tenuipes against Staphylococcus aureus and get the optimal culture conditions. Methods:Single factor test and L9(34)orthogonal design were used to study culture conditions with the diameter of inhibition zone as index. Results:The optimal culture medium was sucrose 4%,yeast powder 2%,MgSO4·7H2O 0.15% and KH2PO4 0.025%. The optimal culture conditions were initial pH 5.5~6.5,media volume 30mL/100mL,inocula amount 5%~7.5% and culture time 6d. Conclusion:The diameter of inhibition zone of Staphylococcus aureus could reach 23.5mm under the conditions.

Ten kinds of Chinese traditional medicines were added into the medium for culturing a fungus Paecilomyces tenuipes.The results showed that the adenosine production in the myceliun and mycelial biomass increased when Redices pueraria,Radix polygoni multiflori and Tartary buckwheat were added.There were increases on mycelial biomass but no change on the adenosine production in the myceliun when Panax notoginseng,Lily and Forsythia suspensa were added.Mycelial biomass decreased when Flos Lonicerae,Chinese wolfberry,Glycyrrhiza and Astragalus membranaceus were added.Based on the results,the effect of initial pH value on the adenosine production in the myceliun was investigated.The results showed that the adenosine production in the myceliun could reach the maximum of 1.984 7 mg/g,when Radix polygoni multiflori was added at 1.25% and the pH value was 7.5.

The effect of oxygen supply on cell growth and nucleoside production in the mycelium was studied in a 30L fermentor,with air saturation at 20%,40%,60% and 80%. The results showed that the maximal biomass was 18.3,21.6,24.7,27.3g/L,respectively,at the 13,11,7,5d and maximal nucleoside production was 3.8643,5.7183,5.2311,4.4968mg/g,respectively,at the 13,13,9,7d. Based on the results,a strategy of two stage DO manipulation was proposed. With such a strategy,production and specific formation rate of nucleosides in the mycelium was 19.9% and 19.2% higher than with one stage manipulation.

On the basis of screening of Paecilomyces tenuipes,Paecilomyces fumosoroseus,Paecilomyces farinosus,Pleurotus ostreatus,Lentinus edodes and Flammulina velutipes,this paper chooses Paecilomyces fumosoroseus as original strain of UV treatment.The cholesterol oxidase producing mutant strain MFEC006 was obtained from Paecilomyces fumosoroseus,the enzyme activity reached 0.548 3 U/mL,154% higher than that of the starting strain.The properties of the mutant strain were stable after eight generations.The optimal fermentation conditions of strain MFEC006 were also studied.The results showed that the optimal culture conditions were as follows: pH 6.5,temperature 27 ℃,inoculum 10% and rotation rate 180 r/min.

采用以麦角甾醇含量为指标间接测定固态发酵产物生物量的方法,对冬虫夏草无性型-中国被毛孢的固态发酵条件进行了研究.正交试验结果表明固态发酵最佳培养基组成为:大米5g,玉米粉2g,蚕蛹粉1g,麸皮2g;单因素试验结果表明:当培养温度20 ℃、料水比1∶ 1.5、料层厚度2 cm、无光照时对菌体生长较为有利.发酵条件优化后,菌丝体中麦角甾醇的含量可达0.5911mg/g,比优化前提高了38.6%.

[Objective] The research aimed to study the effects of ultrasonic extraction on the antioxidant ability of Paecilomyces tenuipes mycelia and confirm the optimum extraction method.[Method] With DPPH free radical scavenging rate as the index,the ultrasonic extraction conditions such as extraction solvent,extraction times,ultrasonic temperature,time,solid-liquid ratio and ultrasonic power were studied by using single factor test and L9(34) orthogonal design. [Result] The optimum extraction process conditions for antioxidant active components from P.tenuipes mycelia by ultrasonic extraction were as followings: with water as extraction solvent,extraction temperature of 30 ℃,extraction time of 30 min,solid-liquid ratio of 1∶30,extraction power of 290 W and extraction times of twice.[Conclusion] Compared with conventional extraction methods,ultrasonic extraction is a kind of simple,time-saving and efficient method.

[Objective] The purpose was to determine the mannitol and scavenging rate of hydroxyl radical in fermented mycilia of Paecilomyces tenuipes by using colorimetry metheod.[Method] With P.tenuipes as tested strains,the mannitol was extracted by using ultrasonic extraction from the mycelia of P.tenuipes.With D-mannitol as the standard substance,the standard curve was drew and the regression equation was calculated.The mannitol content and scavenging rate of hydroxyl radicals were calculated by measuring the Absorbance of mannitol.At the same time,the culture time and extraction time were determined to study their effect on the mannitol content and scavenging rate of hydroxyl radical in mycelium of P.tenuipes.[Result] Mannitol had the maximum Absorption peak at the 412 nm and its linear relationship was better when the content of mannitol was 10-80 μg/ml.The regression equation was C=103.09A-1.381 4,r=0.998 8,and the average recovery was 101.47%,RSD=2.6%(n=5).The trends of the mannitol content and scavenging rate of hydroxyl radical in mycilia did not change with the culture time,and they could reach as highest as 73.56 μg/ml and 47.95% resp.after culture for 6 d.[Conclusion] This study provided a theoretical basis for further study on the antioxidant function of P.tenuipes.

AIM: To investigate the effects of Hirsutella sinensis fermented mycelia on immunologic function in mice. METHODS: The mice were reared with Hirsutella sinensis fermented mycelia at the doses of 50, 200 and 800 mg/kg for 5 days for tests of carbon clearance, serum haemolysin and delayed-type hypersensitivity (DTH) induced by 2-dinitro fluoro benzene (DNFB). RESULTS: The medium and high doses of 200 and 800 mg/kg enhanced phagocytosis by uninucleate macrophages, humoral immunity and cellular immunity of mice significantly with obvious dose dependence. CONCLUSION: These results suggest that the Hirsutella sinensis fermented mycelia has substantial immunoregulation function.

This study analysed the proximate compositions, contents of the bioactive ingredients, amino acids, inorganic elements and fatty acids in the mycelia of Paecilomyces cicadae and fruit body of Cordyceps cicadae. The results showed that in mycelia of Paecilomyces cicadae the content of Cordyceps polysaccharide, mannitol, ergosterol, and adenosine was 33.2 mg/g, 78.9 mg/g, 0.6256 mg/g, and 1.0620 mg/g respectively. The content of microelements such as Ca, Fe, Zn, Se, and Mn was 5187.59 μg/g 207.23 μg/g, 41.93 μg/g and 0.087 μg/g respectively. It was also found that there were 18 kinds of amino acid in the mycelia. In addition , it had a lot of unsaturated fatty acids. Most of them were linoleic acid and octadecenoic acid , which accounted for 53.91% and 20.63% respectively. Compared with Cordyceps cicadae, the contents of Cordyceps polysaccharide, mannitol, ergosterol, adenine, total amino acid, total necessary amino acid and unsaturated fatty acid in the mycelia of Paecilomyces cicadae were higher than those of Cordyceps cicadae. Other chemical compositions were similar to those in Cordyceps cicadae.

The optimum medium and optimal cultivation conditions of Hirsutella sinensis were studied,and the accumulation of nucleosides in the mycelia of shake culture were investigated by ultraviolet spectrophotometry. The results showed that the formula of appropriated culture medium was as follows: glucose 2%,yeast powder 3%,milk powder 1%,MgSO_4.7H_2O 0.1% and KH_2PO_4 0.2%.The suitable culture conditions were as follows: pH 6.5,medium volume 100mL/500mL,rotation rate 180r/rain,inoculum amount 10%.The total nucleoside content reached 6.5748mg/g in the mycelia fermented under optimized culture condition,improved by 33.6% as compared with that under initial culture condition.

The analysis on effective components in the fermentation mycelia and broth of Paecilomyces gunnii var.minor RCEF0864 showed that the contents of aminoacids,raw protein,polysaccharide,ergosterol,adenosine were higher than those in Cordyceps sinensis.Some important metallic elements such as Zn,Fe,Ca.etc.were richer in the fermentation mycelia and broth of P.gunnii var.minor RCEF0864.A DPPH-TLC and DPPH-Mcroplate assay were made to determine the free radical scavenging activity of methanol extract from fermentation mycelia and broth of RCEF0864.The results revealed that the two kinds of extracts has strong free radical scavenging activity.At the concentration of 5.0 mg/mL they can respect a decrease of 53.86% and 59.66% 0.4 mg/mL DPPH radicals after incubated at 37℃ for 10 minutes.

Composition of nucleosides in Cordyceps Sinensis and the fermentation broth of RCEF0273 were determined by high-performance liquid chromatography(HPLC).The results showed that nucleosides in the fermentation broth of RCEF0273 contained mainly adenosine,uridine,guanosine.Nevertheless,the contents of the three nucleosides are many times than those in Cordyceps Sinensis.Therefore,we consider the fermentation broth of RCEF0273 a good resource for medical implication and deserve further research.

研究了金龟子绿僵菌菌株Ma83液体培养最佳营养及环境条件,为无纺布培养提供良好的液体种子.以生物量为指标,通过液体摇瓶试验确定了绿僵菌Ma83菌株液体种子培养参数,对不同的氮源、碳源及其浓度和初始pH进行单因素分析.结果表明,绿僵菌Ma83菌株液体种子培养最佳营养条件为4%黄豆粉,2%白砂糖,0.5%MgSO4·7H2O,0.05g/LKCl,0.1g/L FeSO4·7H2O,1.0g/L KH2PO4,pH6.3～6.5.70L发酵罐培养,干生物量第三天可达35g/L,显著优于筛选前培养基(SDA,28g/L,a=0.05).

Anti Candida albicans disc assay was employed to screen antimicrobial metabolites from the selected entomogenous fungi strains. The result demonstrates that the metabolites of some strains such as Paecilomyces gunii RCEF0866,RCEF0857,Paecilomyces sp. RCEF0189,Baeuveria bassiana RCEF0285 and Nomuraea prasina RCEF0879 have significant anti Candida.albicans activities and their diameter of inhibition zone are 17.28 mm,16.59 mm,17.07 mm,15.66 mm and 22.78 mm,respectively.