来源于枯草杆菌的青霉素G酰基转移酶的生产：培养基成分的影响

分离得到一株产胞内青霉素G酰基转移酶(PAC)的高产枯草杆菌。该菌株中PAC的产生受苯乙酸的诱导。在28℃，pH7．0条件下，考察不同碳源氮源对菌体生长和PAC产生的影响。生长在分别补充蔗糖为碳源和胰蛋白胨为氮源的培养基中的细胞产生的最高活性分别为6．45和8．92u／mg。生长在含有蔗糖和胰蛋白胨培养基中的细胞可产生PAC的最高浓度为10．1u／mg，是基本培养基中的2倍。     

放射型根瘤菌WSH2601生产辅酶Q10的摇瓶发酵条件

以放射型根瘤菌(Rhizobus radiobacterium，WSH2601)作为辅酶Q10的生产菌株。研究了氮源、碳源、接种量、溶氧、初始pH、发酵温度及添加物等因素对细胞生长与产物辅酶Q10合成的影响。结果表明：玉米浆和酶母膏是较好的氮源。葡萄糖与蔗糖是辅酶Q10发酵的较好的碳源。接种量对辅酶Q10发酵的影响不大．为4％。适宜的初始pH值为7，番茄汁能较好地促进细胞的生长。添加玉料浆、L-甲硫氨酸、番茄汁和异戊醇有利于产辅酶Q10；溶氧对细胞生长与产物辅酶Q10合成的影响较显著．通过正交试验初步确定了发酵条件，碳源为1．5g/d1葡萄糖和2．5g／d1蔗糖混合物。酵母膏0．8g／d1．初始pH值7．每500ml装液量为50ml。最后经综合优化条件，在摇瓶发酵条件下：菌体生长量(以干重计)为13．8g／1，发酵液中辅酶Q10产量达到22．9mg／1，比优化前分别提高34％和53％。     

同源四倍体桔梗组培技术的优化研究

为了建立优良同源四倍体桔梗的快速繁殖和离体保存技术，采用均匀设计和正交设计以添加不同激素的MS培养基为基本培养基进行筛选和优化。结果表明：同源四倍体桔梗最佳丛生芽培养基为MS＋BA（6-苄基嘌呤）0．6mg／L＋NAA（萘乙酸）0．4mg／L＋RTCA（病毒唑）3．0mg／L，丛生芽增殖系数可达到9．13。最佳壮苗培养基为：MS＋BA0．2mg／L＋IAA（吲哚乙酸）0．1mg／L＋PP333（多效唑）0．2mg／L＋RTCA3．0mg／L。最佳生根培养基为：1／2MS＋IAA0．7mg／L＋AC（活性炭）0．2mg／L＋ABT（生根粉）0．2mg／L，生根率可达100％。最适室温离体保存条件为：1／4MS＋蔗糖30mg／L＋甘露醇30mg／L＋PP3331．0mg／L＋BA0．3mg／L＋IAA0．1mg／L＋RTCA3．0mg／L，保存150d时存活率仍可达到59．2％，同时恢复生长快，遗传稳定性好。本研究为优良同源四倍体桔梗的推广应用和优良种质离体保存、减少遗传变异提供了技术依据。     

序贯疗法与常规疗法根除幽门螺旋杆菌疗效分析

目的观察由奥美拉唑＋克拉霉素＋左氧氟沙星＋阿奇霉素组成10日序贯疗法根除幽门螺旋杆菌（Hp）的疗效。方法将我院82例确诊为Hp感染阳性的患者分成A组41例和B组41例，A组先给予奥美拉唑20mg，2次／d，＋克拉霉素0．5g，2次／d，5d后给予奥美拉唑20mg，2次／d，＋左氧氟沙星0．2mg，2次／d，＋阿奇霉素0．5g1次／d，疗程5d；B组给予常规奥美拉唑20mg，2次／d＋克拉霉素0．5g，2次／d＋阿莫西林1．0g，2次／d，疗程7d。结果A组根除率94．8％，2例因药物反应不能耐受，6例出现轻微不良反应；B组根除率76．9％，2例因药物反应不能耐受，5例出现轻微不良反应，A组和B组根除率进行z。检验，P〈0．05；两组药物不良反应发生率19．5％、17．0％，P〉0．05。结论以奥美拉唑＋克拉霉素＋左氧氟沙星＋阿奇霉素组成的10日序贯疗法优于常规三联疗法，在不良反应反面无差异。     

GP和NP方案治疗晚期NSCLC的对比研究

目的观察吉西他滨＋顺铂（GP）方案和长春瑞滨＋顺铂（NP）方案治疗非小细胞肺癌的疗效和安全性。方法GP组：GEM1．0～1.2g／m^2第1、8d。DDP25～30mg／m^2第1～3d。NP组：NVB25g／m^2第1、8d，DDP25～30mg／m^2第1～3d。结果GP组和NP组有效率分别为40％和38％。无统计学差异。GP组和NP组中位生存期分别为12．4个月和11．7个月，1年生存率为40％和38．2％，2年生存率为20％和20．1％。无统计学差异。Ⅲ～Ⅳ度白细胞减少发生率GP组和NP组分别为17．2％和28．6％．Ⅲ～Ⅳ度血小板减少率GP组和NP组分别为21．1％和9．5％。均无统计学差异。外周静脉炎发生率GP组和NP组分别为9．6％和42．9％。其中Ⅲ-Ⅳ度发生率GP组和NP组分别为0和12．7％．有统计学差异。其它毒性反应相似。结论GP组和NP组方案治疗晚期非小细胞癌疗效肯定且相近。毒性反应可耐受，可根据临床合理应用。     

奥美拉唑、左氧氟沙星、阿莫西林三联法治疗消化性溃疡的疗效观察

目的评价奥美拉唑、左氧氟沙星、阿莫西林三联法治疗消化性溃疡的疗效。方法85例幽门螺杆菌（Hp）阳性消化性溃疡患者，随机分为治疗组及对照组。治疗组42例，给予奥美拉唑（20mg，2次／d）＋左氧氟沙星（200mg，2次／d）＋阿莫西林（1000mg，2次／d），疗程l周；对照组43例，给予奥美拉唑（20mg，2次／d）＋甲硝唑（400mg，2次／d）＋阿莫西林（1000mg，2次／d），疗程1周。停药4周后复查胃镜，观察Hp根除率、溃疡有效率，并观察症状缓解率及不良反应。结果治疗组Hp根除率为88．1％（37／42），对照组为62．8％（27／43）（χ^2=7．314，P〈0．叭）；治疗组和对照组症状缓解率、溃疡有效率和不良反应发生率分别为92．9％和88．4％、85．7％和72．1％、14．3％和23．3％，两组间差异无统计学意义（P〉0．05）。结论奥美拉唑、左氧氟沙星、阿莫西林三联疗法是治疗Hp阳性消化性溃疡的理想方法之一。     

溶媒耐受性绿脓假单胞菌PseA产生耐溶媒蛋白酶的培养条件优化及产物鉴定

据报道，从源自Pseudomonas aeruginosa的溶媒耐受性绿脓假单胞菌株（PseA）中可获得耐溶媒蛋白酶。本研究所获得产生该酶的优化培养条件：以0．7％甘油为碳源，0．4％酪蛋白和0．6％酵母抽提物作为氮源，pH7．0；种子液培养36h后以2．5％的接种量接种；30℃振荡（250r／min）培养24h。在该条件下，菌体生长最好，酶活力最高（160lu／ml）。该蛋白酶的最适pH与温度为9．0和55℃。     

海南广藿香试管内芽分化与愈伤组织诱导研究

目的选择海南广藿香的最佳芽分化和愈伤组织诱导培养基。方法利用海南广藿香的叶柄、茎尖和带节茎段进行组织培养芽分化试验,并对上述材料进行愈伤组织诱导培养基选择试验。结果在芽分化培养基中,MS＋3-吲哚丁酸（IBA）1．0mg／L＋6-苄基腺嘌呤（6-BA）0．6mg／L培养基能够较好地诱导茎尖和带节茎端长出新芽,诱导率分别为94．3％和94．6％;在愈伤组织诱导培养基中,MS＋萘乙酸（NAA）0．1mg／L＋6-BA2．0mg／L培养基能较好地诱导海南广藿香叶柄和带节茎段产生愈伤组织,诱导率分别为80．0％和100％。结论海南广藿香的最佳芽分化和愈伤组织诱导培养基分别为Ms＋IBA1．0mg／L＋6-BA0．6mg／L和Ms＋NAA0．1mg／L＋6-BA2．0mg／L。     

车前子中槲皮素、木犀草素、山柰酚、芹菜素的含量测定

目的建立车前子的槲皮素、山柰酚、木犀草素、芹菜素含量测定方法。方法采用反相高效液相色谱（RP—HPLC）法,色谱柱为Zorbax SB—C18柱（150mm×4．6mm,5μm）;检测波长360nm,流动相为甲醇-0．2％磷酸（45：55）,柱温30℃,流速1．0mL／min。结果槲皮素、山柰酚、木犀草素、芹菜素的回归方程分别为Y=1504．412X＋9．9756,Y=1991．745X＋8．6051,Y=567．591X＋2．5397,Y=1811．803X＋0．3074;质量浓度线性范围分别是0．05522～0．1933μg／mL,0．0461—0．1613μg／mL,0．1255～0．4393μg／mL,0．0603～0．211μg／mL,r在0．9999．1．0000之间;槲皮素、山柰酚的加样回收率分别为92．87％和102．95％,RSD分别为0．28％和1．08％;样品分别含槲皮素、山柰酚、木犀草素、芹菜素0．81,0．634,0,0．08mg／g。结论RP—HPLC法分析车前子中黄酮化合物的含量,方法简便可行,重复性好,结果可靠。     

高效液相色谱法测定西洋参冲剂中人参皂甙Rb_1的含量

采用固相萃取净化技术及高效液相色谱法，测定了西洋参冲剂中人参皂甙Rb1的含量，结果在12.5～200μg／ml范围内，Rb1的含量与峰面积呈良好线性，平均回收率及RSD分别为：101．3±3．2％（0．4mg，n＝5）；101．2±2．6％（0．2mg，n＝5）；99．0±4．4％（0．1mg，n＝5）。最小检出量为0．3μg，本法简便、快速、准确。     

Enhanced sinefungin production by medium improvement, mutagenesis and protoplast regeneration of Streptomyces incarnatus NRRL 8089

Increased production of sinefungin, a very potent antifungal and antiparasitic nucleoside antibiotic was achieved by medium and strain improvement. When soybean-meal, dextrin and yeast extract were added as carbon and nitrogen sources to the fermentation medium, instead of corn steep liquor, soya-oil and glucose; the antibiotic yield increased from 40 micrograms/ml to 126 micrograms/ml with low biomass production. Strain improvement was attempted by two methods. The mean antibiotic yield of the variants after multistep mutagenesis by N-methyl-N'-nitro-N-nitrosoguanidine and ethyleneimine was 466 micrograms/ml. Protoplasts of the parental strain were prepared by lysozyme digestion from mycelia grown in a medium containing 0.7% glycine. The mean activity of the regenerated protoplasts was 664 micrograms/ml. Thus, the overall sinefungin production could be increased 16-fold.     

天然与发酵培养冬虫夏草中核苷类成分的含量及其变化

目的 比较天然和人工冬虫夏草中核苷类成分的含量,并对其变化进行考察。方法 采用高效毛细管电泳法测定天然和人工冬虫夏草中核苷类成分的含量;考察湿热对冬虫夏草中核苷类成分含量的影响。结果 人工冬虫夏草的核苷含量明显高于天然冬虫夏草;新采集的天然冬虫夏草中核苷类物质含量极低,而采集日久者核苷类成分含量却较高;湿热可明显增加天然冬虫夏草中核苷类物质的含量,但对人工冬虫夏草中核苷类成分的含量无明显影响。结论 天然和人工冬虫夏草中核苷类成分存在一定差异,天然冬虫夏草中核苷类物质可能主要来源于大分子核苷(酸)的降解,腺苷在天然冬虫夏草质量控制中的应用价值尚值得进一步研究。     

Preparation of culinary-medicinal maitake mushroom, Grifola frondosa (Dicks.: Fr.) S.F. Gray (Aphyllophoromycetideae)-fermented wheat and its antioxidant properties

The optimization of solid-state fermentation conditions for mycelial growth in wheat by culinary-medicinal maitake mushroom, Grifola frondosa (Dicks.: Fr.) S.F. Gray was studied. The results from the center of the design analysis showed that the recommended optimal conditions for carbon, nitrogen sources, and temperature were 19 mg lactose/g, 2.0 mg malt extract/g, and 30 degrees C, respectively. Under these conditions, the mycelial biomass of 533.1 mg/g was obtained in fermented wheat, comparable to that (545.7 mg/g) obtained using the one-factor-at-a-time method. Based on the results obtained, EC50 values of ethanolic and hot water extracts from fermented wheat were 1.13-15.27 mg/mL, indicating that fermented wheat was effective in antioxidant activity, reducing power, scavenging ability on 1,1-diphenyl-2-picrylhydrazyl radicals, and chelating ability on ferrous ions. Total phenol contents of both extracts were 13.35 and 23.01 mg/g, respectively. Accordingly, the information obtained would be applicable for the cultivation and consumption of G. frondosa-fermented wheat to provide its beneficial health effects.     

天然与发酵培养冬虫夏草中核苷类成分的含量及其变化

目的比较天然和人工培养冬虫夏草中核苷类成分的含量,并考察其变化.方法用高效毛细管电泳法测定天然和人工冬虫夏草中核苷类成分的含量;考察湿热对冬虫夏草中核苷类成分含量的影响.结果人工冬虫夏草的核苷含量明显高于天然冬虫夏草;新采集的天然冬虫夏草中核苷类物质含量极低,而采集日久者核苷类成分含量较高;湿热可明显增加天然冬虫夏草中核苷类物质的含量,但对人工冬虫夏草中核苷类成分的含量无明显影响.结论天然和人工冬虫夏草中核苷类成分存在一定差异,天然冬虫夏草中核苷类物质可能主要来源于大分子核苷(酸)的降解.     

Medium optimization for enhanced co-production of two bioactive metabolites in the same fermentation by a statistical approach

This paper describes improved optimization method that combines the one-factor-at-a-time method (OFAT), Plackett-Burman design, and the response surface method (RSM), which were used to optimize the medium for the production of fumigaclavine C (FC) and helvolic acid (HA) from endophytic Aspergillus fumigatus CY018 simultaneously. The ideal carbon and nitrogen sources for the two compounds were assessed initially via the one-factor-at-a-time method. Three key cultivation factors (pH, phosphate, and inoculum size) were chosen based on the results of Plackett-Burman design, and subsequently optimized by the central composite design. The two metabolites were amply afforded when the cultivation was carried out with the inoculum size of 2.45% at pH 4.2 and 28°C for 19 days in the medium containing (g/l): mannitol 50, sodium succinate 5.4, NaNO? 2, MgSO?·7H?O 0.3, FeSO?·7H?O 0.01, and KH?PO? 0.67. The highest yields of FC and HA achieved herein were 17.26 and 16.88 mg/l. This work might be the first endeavor leading to the improved simultaneous production of two complex active metabolites with a single strain.     

竹黄多糖发酵工艺的优化研究

目的:优化竹黄多糖的发酵工艺。方法:培养竹黄菌菌丝体液,以菌丝干重和多糖得率为指标,分析竹黄多糖发酵过程中碳源、氮源、pH值对多糖积累的影响。结果:蔗糖、玉米粉、酵母膏均有利于菌丝的生长和多糖的形成。通过正交试验确定了竹黄多糖最佳发酵培养基:蔗糖3%、玉米粉3%、酵母膏0.5%、麸皮2%、(NH4)2SO40.2%、MgSO4.7H2O0.05%、KH2PO40.1%;在10L发酵罐上竹黄多糖含量最高达10g.L-1。结论:该工艺稳定,可为工业生产提供理论依据。     

Optimization of fermentation process of Cordyceps militaris and antitumor activities of polysaccharides in vitro

The influence of medium composition and cultural conditions on simultaneous yield of mycelia, intracellular polysaccharide, adenosine, and mannitol by Cordyceps militaris CGMCC 2909 was investigated with desirability functions in this study. An optimization strategy based on the desirability function approach, together with response surface methodology (RSM) has been used to optimize medium composition, and the optimal medium was obtained via the desirability as follows: yeast extract 10.33 g/L, sucrose 27.24 g/L, KH₂PO₄ 5.60 g/L and the optimal culture conditions are initial pH 6, 25°C, rotation speed 150 r/minute, inoculum size 4%(v/v), and medium capacity 40 mL/250 mL. Under these conditions, the yield of mycelia, intracellular polysaccharide, adenosine and mannitol reached 12.19 g/L, 0.6 g/L, 61.84 mg/L, and 1.38 g/L, respectively, and the D value was 0.77. Furthermore, the polysaccharides showed significant antitumor activities against HeLa and HepG2 in vitro in a dose-dependent manner in 72 hours. At a concentration of 1000 mg/mL, the inhibition rate of polysaccharides was 92.38% and 98.79%. The IC50 for HeLa and HepG2 were 70.91 μg/mL and 97.63 μg/mL, respectively.     

糖肽类抗生素A40926B产生菌的培养基优化及菌种选育

以A40926 B产生菌野野村放线菌SIPI-D-30为出发菌株,通过UV诱变及乙酸钠和甘氨酸抗性平板筛选,获得A40926 B产量提高的突变株SIPI-U-157。经优化,确定摇瓶发酵培养基(g/L)为:葡萄糖5.0、麦芽糊精30.0、可溶性淀粉10.0、玉米淀粉30.0、大豆蛋白胨5.0、肉蛋白胨10.0、酵母浸粉5.0、冷榨黄豆饼粉10.0、棉籽饼粉5.0、L-缬氨酸1.0,发酵培养7 d,突变株的A40926 B发酵单位为893 mg/L。该突变株在5 L发酵罐中培养180 h,A40926 B的产量最高为583 mg/L。     

Medium optimization for enhanced co-production of two bioactive metabolites in the same fermentation by a statistical approach

This paper describes improved optimization method that combines the one-factor-at-a-time method (OFAT), Plackett–Burman design, and the response surface method (RSM), which were used to optimize the medium for the production of fumigaclavine C (FC) and helvolic acid (HA) from endophytic Aspergillus fumigatus CY018 simultaneously. The ideal carbon and nitrogen sources for the two compounds were assessed initially via the one-factor-at-a-time method. Three key cultivation factors (pH, phosphate, and inoculum size) were chosen based on the results of Plackett–Burman design, and subsequently optimized by the central composite design. The two metabolites were amply afforded when the cultivation was carried out with the inoculum size of 2.45% at pH 4.2 and 28°C for 19 days in the medium containing (g/l): mannitol 50, sodium succinate 5.4, NaNO₃ 2, MgSO₄·7H₂O 0.3, FeSO₄·7H₂O 0.01, and KH₂PO₄ 0.67. The highest yields of FC and HA achieved herein were 17.26 and 16.88 mg/l. This work might be the first endeavor leading to the improved simultaneous production of two complex active metabolites with a single strain.     

Production of biomass, polysaccharides, and ganoderic acid using non-conventional carbon sources under submerged culture of the Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W.Curt.:Fr.)P. Karst. (higher Basidiomycetes)

The effect of different non-conventional carbon sources was studied in the submerged culture of Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum, for simultaneous production of mycelial biomass, bioactive ganoderic acid, and polysaccharides, in less time, using non-conventional carbon sources to minimize the high costs of current culture media. The optimal medium composition was defined as (g/L): 50 of barley flour, 0.2 of KH2PO4, 0.1 of MgSO4?7H2O, and 1 NH4Cl. Cultivated under this complex culture medium, the mycelial biomass production was 23.49 ± 0.37 g/L; the extracellular polysaccharides production was 2.72 ± 0.11 g/L; the intracellular polysaccharides production was 2.22 ± 0.06 g/L; the ganoderic acids production was 299.67 ± 11.63 mg/L. One liter of culture medium developed in this project was priced at USD $ 0.11 if barley flour is used as carbon source or $ 0.13 with oat flour in order to get a good amount of products of interest.