Kluyveromyces fragilis LFS-8611 β-D-半乳糖苷酶的摇瓶培养条件

脆壁克鲁维酵母(Kluyveromycesfragilis)LFS 8611合成的β D 半乳糖苷酶具有较高的催化半乳糖基转移反应活力.脆壁克鲁维酵母(K.fragilis)LFS 8611细胞生长和β D 半乳糖苷酶的合成同步.该菌株生长和产酶的最适碳源为半乳糖,乳糖次之;最适氮源为蛋白胨F403;最适培养条件为:发酵培养基的初始pH值为7.0,摇床的转速为200r/min.培养基中碳源和氮源质量浓度对菌体生物量和β D 半乳糖苷酶活力有重要影响,以12mg/mL乳糖为碳源,16mg/mL蛋白胨(F403)为氮源,在最适培养条件下培养32h后,菌体生物量和β D 半乳糖苷酶活力分别为7.56g/L和18.83U/mL.     

正红菇液体发酵培养基和发酵条件的研究

研究了正红菇(Russulavinosa)在发酵过程中菌丝体及胞外多糖产量的变化趋势,碳源、氮源、无机盐以及发酵温度、pH值、时间和装液量等因素对正红菇液体深层发酵菌丝产量的影响。结果表明,蔗糖为最佳碳源,酵母膏为最佳氮源,优化培养基配方为蔗糖40g/L,酵母膏9g/L,KH2PO42g/L,MgSO41g/L;最适菌丝体生长的液体发酵条件:培养温度28℃￣30℃,初始pH值为5.5￣6.5,250mL三角瓶装液量为50mL￣60mL,发酵时间为5d。通过优化培养基和发酵条件,胞外多糖产量达到4.96g/L,菌丝体生物量达到22.34g/L。     

金针菇液体培养营养需求及培养条件研究

在装液量100mL/250mL三角瓶,接种量10%的基础上,以菌丝体生物量为指标,从实验室常见的5种碳源、6种氮源中,筛选出金针菇菌丝体生长的最适碳源、最适氮源和最适pH,并进一步通过正交实验优化出培养基的养分浓度和摇瓶发酵条件,从而确定金针菇摇瓶培养基组成为葡萄糖25g/L,NH4NO31.5g/L,KH2PO41.0g/L,MgSO4·7H2O0.5g/L,VB150mg/L;最佳发酵条件为23℃,起始pH6.0,摇床转速150r/min,发酵时间8d。     

1株高产EPS嗜热链球菌的筛选及培养条件优化

为了研究乳酸菌高产EPS的条件,以产EPS量为指标,从8种复合菌株中筛选出1株嗜热链球菌,通过单因素和正交试验对该菌EPS合成培养基中的碳源、氮源、培养条件进行了研究。结果显示:该菌在优化的培养基中EPS生成量可达到145.48mg/L;合成胞外多糖最适培养条件为:接种量2%,初始pH6.5,发酵温度34℃,发酵时间36h。在此条件下EPS生产量可高达270mg/L,提高了116%。因此氮源组合和pH值为影响EPS合成的显著因素(p<0.05)。在恒定pH值条件下发酵培养可以提高EPS的生成量,即474mg/L,又提高了72%。     

假单胞菌L-3对氧化乐果降解条件的优化

为确定氧化乐果降解菌假单胞菌L-3最佳的降解培养基配比及最适的环境条件,在含一定质量浓度氧化乐果的分离培养基中,对在不同碳源、氮源和磷源条件下的培养基进行降解实验,并通过正交旋转组合实验优化设计,优选出最适的碳源、氮源和磷源及其质量浓度;对菌株在不同的温度、pH值、接种体积分数、初始氧化乐果质量浓度环境条件下进行降解,通过测定降解率优化出最适的环境条件。结果表明,菌株L-3最适降解条件为:蔗糖1.7 g/dL,牛肉蛋白胨0.92 g/dL,KH2PO40.184 g/dL;最佳的降解环境条件为pH 7、30℃、接种体积分数3%、初始氧化乐果质量浓度为50 mg/L;优化后对氧化乐果的降解率高达82.59%。     

柏树火焰层孔菌菌丝体发酵工艺条件优化

柏树火焰层孔菌是中国以及亚洲菌物的新记录属中的新记录种,是一种稀有的药用菌。为寻求高产的生长条件和经济的培养基,进行转速、温度、pH 值、培养液装瓶体积、接种量、碳源、氮源、C/N 对菌丝体生长影响的单因素试验和多因素试验设计,利用液体培养基探索这些条件对菌丝体产量的影响。结果表明：菌丝 生长最适碳源为淀粉,最适氮源为麸皮,P、K、Mg 对菌丝的生长具有促进作用,最适C/N 为20:1～25:1;菌丝生长的最适转速为150r/min,温度为25～30℃,最佳pH 值为5～7,最适培养液装瓶体积为100mL,最适接种量为8mL/100mL。液体培养基的最佳营养条件为：葡萄糖 25g/L、麸皮 125g/L、 K2HPO4 0.25g/L、pH 6.5。通过生长曲线实验得到Logsitic 方程为：y = 0.22/(1+ 1.019e － 0.013x)。     

木霉T68发酵基质的研究

以生物量(孢子量和菌丝体)为指标,菌落直径为参考,采用单因素和正交试验对木霉T68(Trichoderma"T68")的发酵条件进行优化.优化后的木霉发酵控制参数为固体培养最佳组合为pH7、30℃、7d,碳源为蔗糖,氮源为丙氨酸.最适T68菌丝生长的液体培养基是以玉米粉和麸皮分别作为碳、氮源,最佳培养基配方为麦麸30g/L,玉米粉30g/L,葡萄糖7.5g/L,KH2PO41g/L;发酵培养时间为3d;培养基的初始pH值为5.0～7.0;发酵温度为(30±1)℃;摇床转速为160r/min.最佳培养条件下菌丝干重为3.272g/L.     

灵芝液体发酵条件优化研究

对灵芝液体发酵温度、发酵时间和灵芝液体发酵培养基的氮源、碳源及其最适浓度、初始pH等发酵条件进行了研究，结果表明，灵芝液体最适发酵条件为：氮、碳源分别为蛋白胨、葡萄糖，碳源浓度为3％，培养基初始pH为6．5，发酵温度为28℃，发酵时间为5d。     

胞苷的发酵培养基优化

为了提高大肠杆菌生产胞苷的能力,研究了培养基中碳氮源对菌株生产胞苷的影响,从而确定发酵培养基的最适碳源、氮源,并同时确定其最适使用浓度;经实验验证,发酵培养基中的最适碳源为甘油,其使用浓度为80g/1;最适氮源为酵母膏和蛋白胨,其使用浓度分别为20g/l和5g/l.在优化的培养基条件下,胞苷积累量相较于初始培养基提高了57.9％.     

产葡萄糖氧化酶黑曲霉紫外诱变及产酶条件的研究

目的提高黑曲霉中葡萄糖氧化酶的产量。方法紫外诱变法选出产酶优势菌株,优化碳源、氮源、碳酸钙、发酵时间等条件。结果黑曲霉发酵液酶活达到6.5 U/mL,比初始酶活提高5倍;单因素条件试验表明,最适碳源是蔗糖,最适氮源是蛋白胨和NaNO₃,发酵周期为48 h,培养温度是30℃,液体培养基的初始pH值为6.0产酶效果最好。结论紫外诱变后,葡萄糖氧化酶的活力有明显提高。     

甜叶菊毛状根的诱导培养及茉莉酸甲酯对其绿原酸类物质积累的影响

以发根农杆菌ACCC10060诱导甜叶菊毛状根,建立毛状根培养生产绿原酸类物质体系,研究茉莉酸甲酯对绿原酸类化合物积累的影响。经发根农杆菌ACCC10060侵染的甜叶菊叶片外植体,共培养14 d后产生毛状根。聚合酶链式反应检测结果表明,发根农杆菌Ri质粒的rol B和rol C基因已成功整合到甜叶菊毛状根基因组中。MS液体培养基较B5、WPM更利于甜叶菊毛状根生长及绿原酸类物质的积累,培养35 d后,干质量增加约30倍,绿原酸、3,5-二咖啡酰奎宁酸、4,5-二咖啡酰奎宁酸最高含量分别为3.47、11.47、3.04 mg/g。毛状根在MS液体培养基中培养至第3周时分别添加15、45、100μmol/L的茉莉酸甲酯进行诱导,处理后第1、2、4、8天收获,毛状根的生长量和绿原酸类物质含量均有提高,其中以45μmol/L茉莉酸甲酯的促进作用最为显著,3种绿原酸类物质的总产量是对照组的2.68倍(P0.01)。以上结果表明,甜叶菊毛状根培养可用于绿原酸类物质的生产,经茉莉酸甲酯处理可显著提高绿原酸类物质的产量。     

果蔬生防菌葡萄有孢汉逊酵母CM2糖蜜培养基的优化

以葡萄有孢汉逊酵母CM2为研究对象,细胞生物量为测定指标,利用甘蔗糖蜜作为发酵原料,对糖蜜培养基的优化进行较为全面的探讨。采用煮沸法、磷酸法、硫酸法、活性炭法、亚铁氰化钾法对糖蜜进行预处理,结果表明硫酸处理法优于其他处理方法。通过单因素试验研究糖蜜质量浓度、氮源及其添加量、初始pH值、磷酸盐、硫酸镁、氯化钠等培养基组分对CM2生长的影响;采用Plackett-Burman试验设计、最陡爬坡试验和中心组合试验设计对培养基组分进行优化,得到优化培养基组分：预处理后糖蜜78.9 g/L、酵母粉2.2 g/L、初始pH 6.0。通过验证实验获知与PDB培养基相比,优化后糖蜜培养基使CM2生物量提高了83.1%。     

青钱柳悬浮细胞的培养及其基质消耗的规律

青钱柳为我国特有的胡桃科青钱柳属单种属珍稀植物,也是国家重点保护的濒危植物之一,青钱柳叶含三萜、黄酮、多糖及微量元素等活性成分,具有降糖、调脂、增强免疫等多种生理活性和药理功能,是很好的天然保健食品资源。本文建立青钱柳细胞悬浮培养体系,测定悬浮培养细胞的生长曲线,并研究了培养代数、接种量、装液量、pH值和激素组合对青钱柳悬浮体系建立的影响,同时研究了悬浮培养青钱柳细胞对培养液中主要基质的消耗规律。结果表明:适合青钱柳悬浮培养的培养基为:MS+0.1 mg/L2,4-D+0.5 mg/L NAA+1 mg/L KT+30 g/L蔗糖,接种量为8%,摇瓶装液量的体积分数为40%,初始pH 5。青钱柳细胞生长与碳源、氮源和磷的消耗存在着密切关系,在培养8 d,糖和PO43-基本消耗完毕;在氮源的吸收上,NH4+优于NO3-,NH4+的吸收基本上是呈直线下降。     

ENHANCEMENT OF SECONDARY METABOLITE PRODUCTION IN HAIRY ROOT CULTURES OF BETA VULGARIS AND TAGETES PATULA UNDER THE INFLUENCE OF MICROALGAL ELICITORS

The effect of aqueous extracts of green alga-Haematococcus pluvialis and blue green alga-Spirulina platensis was studied for accumulation of betalaines (food colourant) and thiophenes (insecticide useful against food grain spoilage) in hairy root cultures of Beta vulgaris and Tagetes patula, respectively. It was found that upon treatment with extracts (100 μg dry cell extract/ml of the culture medium each) of H. pluvialis and S. platensis there was an increase in biomass of cultured Beta vulgaris hairy roots which was 165.3 g fresh wt/L (14.38 g dry wt/L) and 149.4 g fresh wt/L (13 g dry wt/L), respectively from the initial inoculum of 1.25 g fresh wt/L (0.115 g dry wt/L). The accumulation of betalaines showed an increase of 2.28 fold on 15th day in hairy root culture treated with H. pluvialis over the control (7.9 mg/g dry wt) on the same day from the initial level of 0.67 mg/g dry wt. Similarly S. platensis extract treated hairy roots showed elicitation of betalaines 1.16 fold on 25th day over the control (11.08 mg/g dry wt) from the initial level of 0.67 mg/g dry wt. It was also observed that the H. pluvialis extract also influenced the biomass production in T. patula hairy roots which was 1.4 fold higher than the control (40 g fresh wt/L or 3.21 g dry wt/L) on 20th day from the initial inoculum of 1.25 g fresh wt/L (0.105 g dry wt/L). Whereas the accumulation of thiophene increased by 1.2 fold in cultures treated with H. pluvialis extract on the 20th day over the untreated control (510 μg/g dry wt) from the initial level of 200 μg/g dry wt, while S. platensis extract did not influence the growth as well as thiophene accumulation. In conclusion S. platensis extract influenced elicitation of betalaines in B. vulgaris hairy roots where as H. pluvialis extract elicited betalaine and thiophene production in hairy root cultures of B. vulgaris and T. patula respectively.     

紫色红曲霉FBKL3.0018液态发酵产红曲色素条件的优化研究

以分离自贵州某浓香型酒厂中温大曲的紫色红曲霉（Monascus purpureus）FBKL3.0018为研究对象,以发酵液中红曲色素色价为考察指标,对紫色红曲霉FBKL3.0018产红曲色素的发酵培养基配方进行优化。考察了碳源、氮源、无机盐、生长因子及初始pH值对红曲色素生产的影响,选取对红曲色素生产影响较显著的蛋白胨、FeSO4和初始pH进行响应面优化试验。得到最佳培养基配方为：葡萄糖60 g/L、蛋白胨26 g/L、FeSO4 0.9 g/L、L-谷氨酸 2 g/L和初始pH 4.5。在此优化条件下,红曲色素色价为105.22 U/mL,比优化之前（33.62 U/mL）提高了3.13倍。同时,通过验证试验,实际值105.22 U/mL与预测值108.82 U/mL相对误差为0.97%,说明所建立的回归模型可靠。     

Enhancement of growth and coumarin production in hairy root cultures of witloof chicory (Cichorium intybus L.cv. Lucknow local) under the influence of fungal elicitors

Studies were conducted to determine the possible production of coumarins in hairy root cultures of Cichorium intybus L.cv.Lucknow local under the influence of microbial agents. Pythium aphanidermatum and Phytopthora parasitica var. nicotiana were cultured and their mycelial and medium filtrate were used for the elicitation of coumarin production. The media filtrate of P. parasitica at 1.0% v/v added to the MS basal medium led to the maximum growth of hairy roots of C. intybus, which was 1.57-fold higher than the untreated control on the 28th day of culture, along with maximum esculin and esculetin yields which were 4.06- and 3.71-fold higher than the control on the 28th day. The yields of esculin and esculetin in hairy root cultures of chicory strongly correlated with growth. To check the effect of these fungal elicitors on endogenous polyamine metabolism, titers of total endogenous polyamines were analyzed. It was confirmed that the media filtrate of P. parasitica at 1.0% v/v concentration resulted in maximum accumulation of total endogenous polyamines, wherein endogenous spermine titers were found to be maximum as compared to endogenous spermidine and putrescine titers on the 28th day. Total endogenous spermine in the case of 1.0% MF v/v P. parasitica was 1.3-fold higher than that of the control on the 28th day of culture. Maximum growth index with greater length of primary root (17.61+/-0.18 cm) and greater number of secondary and tertiary roots was recorded for the medium filtrate of P. parasitica at 1.0% v/v concentration. This study also provided an insight into the morphological changes in terms of branching patterns, occurring in roots under the influence of these fungal elicitors.     

Torulopsis sp.ERY237产赤藓糖醇工艺条件的研究

以Torulopsis sp.ERY237作为出发菌株,考察了不同碳源、氮源、无机盐类以及温度等因素对菌种产赤藓糖醇的影响,建立和优化了赤藓糖醇摇瓶发酵培养基配方、发酵工艺条件,同时研究了发酵过程中菌体生物量、pH值、产物浓度的动态变化。结果表明,菌株的最适培养基配方为(g/L):葡萄糖300,玉米浆3.5,C_([Cu~(2+)])1.5,C_([Mn~(2+)])10;适宜的培养条件为初始pH值自然,温度30℃,装液量50 mL/500 mL,转速200 r/min,在此条件下培养132 h赤藓糖醇产量达87.8 g/L,是优化前产量的1.9倍,发酵时间缩短了12 h。     

响应面法优化豆乳链球菌增殖培养基

针对优良酸豆乳发酵菌株豆乳链球菌（Streptococcus sojalactis）SY1.1增殖培养基进行优化,研究碳氮源对豆乳链球菌生长的影响,并采用响应面法优化豆乳链球菌SY1.1的增殖培养基配方。结果表明：以AST为基础培养基,优选大豆蛋白胨和蔗糖为最适氮源和碳源,质量比为1∶2,采用Plackett-Burman设计对11 种促乳酸菌生长因子进行评价,利用中心旋转组合设计和响应面分析确定增殖培养基的配方为：蔗糖10 g/L、大豆蛋白胨20 g/L、磷酸氢二钾2 g/L、番茄汁85.8 mL/L、胡萝卜汁109 mL/L、玉米浆6 g/L,pH 6.8。SY1.1以接种量2%,在37 ℃培养12 h时,活菌数可达（8.9±0.2）×108 CFU/mL,较初始豆浆培养基的活菌数（1.03×108 CFU/mL）提高了7.64 倍。