The influence of phenylalanine on accumulation of rosmarinic and caffeic acids by Lavandula vera MM cell culture

Lavandula vera MM cell suspension culture was grown in Linsmayer-Skoog medium with different concentrations of phenylalanine. Adding phenylalanine to the medium (0.1–0.5 g/l) enhanced accumulation of caffeic acid in parallel with rosmarinic acid. When 0.3 g phenylalanine/l was added, the yield of rosmarinic and caffeic acids reached 87 mg/l and 60 mg/l respectively, compared with 68 mg/l and 4 mg/l in controls (without phenylalanine).     

Rosmarinic acid production by Lavandula vera MM cell-suspension culture

The time courses of growth and rosmarinic acid production by Lavandula vera MM cell suspension were investigated. The uptake of the main nutrients (sucrose, nitrogen, phosphorus, K, Ca, Mg) was followed during cultivation and the data on the physiology of the L. vera MM cell culture are presented. It was established that the cell culture synthesizes rosmarinic acid during the linear phase of growth for a relatively short period (between the 4th and 8th days of cultivation). The influence of sucrose concentration in the nutrient medium on cell growth and accumulation of rosmarinic acid by L. vera MM cell culture was investigated. The results showed that 7% sucrose in the nutrient medium ensured a steady growth of the cell suspension and increased the yield of rosmarinic acid (29.2 g/l dry biomass and 507.5 mg/l rosmarinic acid compared to 13.0 g/l dry biomass and 68.6 mg/l rosmarinic acid for the control cultivation with 3% sucrose). Received: 17 September 1996 / Received revision: 31 January 1997 / Accepted: 1 February 1997     

Production of rosmarinic acid by<Emphasis Type="Italic">Lavandula vera</Emphasis> MM cell suspension in bioreactor: effect of dissolved oxygen concentration and agitation

The relationship between dissolved oxygen (DO) concentration, agitation rate and growth of Lavandula vera MM and rosmarinic acid biosynthesis was investigated in 3 l laboratory bioreactor. Lavandula vera MM cell suspension accumulated the highest amounts of biomass (34.8 g/l) and rosmarinic acid (1870.6 mg/l) on day 12 of cultivation at 50% dissolved oxygen and agitation speed 100 rpm and at 30% dissolved oxygen and agitation speed 300 rpm, respectively.     

Rosmarinic acid by Lavandula vera MM cell suspension: Phosphorus effect

Summary The growth of Lavandula vera MM cell suspension and biosynthesis of rosmarinic acid were followed during its cultivation in Linsmayer - Skoog (LS) medium (containing 170 mg/L KH₂PO₄, control cultivation) and in modified LS media, containing different concentration of KH₂PO₄. Doubled concentration of KH₂PO₄ (340 mg/L) caused an increase in the amount of biosynthesised cell biomass (17 g/L vs 13 g/L for control cultivation) and rosmarinic acid (140 mg/L vs 68 mg/L for control cultivation).     

Enhanced rosmarinic acid production by Lavandula vera MM cell suspension culture through elicitation with vanadyl sulfate

The influence of elicitation on rosmarinic acid biosynthesis by Lavandula vera MM cell suspension culture was investigated using vanadyl sulfate as an abiotic elicitor. It was established that 12 h after treatment with 25 mg/l vanadyl sulfate the rosmarinic acid production was increased up to 3.92 g/l (2.8 times higher compared to the control cultivation). No significant amounts of rosmarinic acid were detected in the culture medium in comparison with its intracellular content. However, it was observed that the extracellular content of rosmarinic acid is 3.3 times higher compared to the control variant (4 h after treatment at elicitor concentration 25 mg/l).     

Radical quenching by rosmarinic acid from Lavandula vera MM cell culture

This study was conducted to evaluate the radical scavenging capacities of extracts and preparations from a Lavandula vera MM plant cell culture with different rosmarinic acid content and to compare them with pure rosmarinic and caffeic acids as well. The methods, which were used are superoxide anion and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radicals scavenging assays. Results showed that extracts and preparations from Lavandula vera MM possess strong radical scavengers, as the best both radical scavengers appeared to be the fractions with enriched rosmarinic acid content, obtained after ethylacetate fractioning (47.7% inhibition of superoxide radicals and 14.2 microM 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid equivalents, respectively). These data reveal the possibilities for application of these preparations as antioxidants.     

Rosmarinic acid production by Lavandula vera MM cell suspension: the effect of temperature

Lavandula vera MM cell suspension, grown at 28 degrees C in a 3-l bioreactor, produced rosmarinic acid maximally at 3 g l(-1)) though most biomass (33.2 g dry wt l(-1)) was at 30 degrees C.     

Rosmarinic acid production by Lavandula vera MM cell suspension culture: nitrogen effect

The growth of Lavandula vera MM cell suspension and the biosynthesis of rosmarinic acid (RA) were followed during its cultivation in Linsmayer–Skoog media, containing different concentrations of ammonium and nitrate ions. The results showed that cultivation in a medium with 0.09g ammonium ions/l (1/4 of standard medium) ensured intensive growth (16g dry biomass/l) and enhanced biosynthesis of RA (15mg/g dry biomass). Cultivation of L. vera MM cell culture in a medium with 1.2-fold concentration of nitrate ions led to accumulation of 11mg RA/g dry biomass which was twice as much as in the standard Linsmayer–Skoog medium.     

Elicitation of rosmarinic acid by <Emphasis Type="Italic">Lavandula vera</Emphasis> MM cell suspension culture with abiotic elicitors

The growth of Lavandula vera MM plant cell suspension culture and rosmarinic acid biosynthesis under elicitation with benzothiadiazole and methyl jasmonate were investigated. Upon elicitation with 50 μM methyl jasmonate, the production of rosmarinic acid was enhanced 2.4-fold (3348 mg/l) compared to the non-elicited cells. The influence of benzothiadiazole on rosmarinic acid biosynthesis was weaker and 12 h after its addition the achieved yields were 20–30% higher compared to the control variant at this time. The influence of both elicitors on rosmarinic acid secretion into the culture medium was also discussed.     

Release of rosmarinic acid by Lavandula vera MM cell suspension in two-phase culture systems

Studies were conducted on the cultivation of Lavandula vera MM cell suspensions in different culture systems for the release of extracellular rosmarinic acid (RA). It was established that during cultivation with Amberlite XAD-4 as a second phase, 6.4% of the total content of RA was adsorbed. When L. vera MM cell suspension was cultivated in an aqueous two-phase system formed by adding 4% polyethylene glycol (MW 20,000) and 7.5% dextran (MW 70,000), 11.8% of the total RA content was released into the top polyethylene glycol phase.     

海洋红酵母Y2发酵产类胡萝卜素条件的研究

目的 对海洋红酵母Y2高产类胡萝卜素的发酵条件进行优化.方法 在摇瓶条件下,研究培养基成分和培养条件对海洋红酵母Y2生长和类胡萝卜素合成的影响,同时进行海洋红酵母Y2发酵过程的动态分析.结果 海洋红酵母Y2优化培养基组合为葡萄糖45 g/L,蔗糖15 g/L,酵母粉5 g/L,蛋白胨2.5 g/L,磷酸二氢钾1 g/L,磷酸二氢钠3 g/L,硫酸镁7.5 g/L,氯化钾3 g/L,氯化钠5 g/L.最适培养参数为:温度20℃,培养基初始pH为5,接种量为10％,250 mL摇瓶装液量为10～50 mL.类胡萝卜素的合成主要集中在对数生长期和稳定期.海洋红酵母Y2最适收获时间为72 h.种龄以36 h为宜.结论 利用优化培养基,在最适条件下培养海洋红酵母Y2,类胡萝卜素产量达到4.97 mg/L,比基础培养基提高了60.32％.     

拉曼被孢霉产γ-亚麻酸的发酵条件优化

对影响发酵产γ-亚麻酸的因素（温度、N源、P源等）进行单因素考察。结果发现,各因素对拉曼被孢霉（Mortierella ramanniana NBRC 8187）产γ-亚麻酸均有不同程度的影响。通过正交试验设计对KH2PO4、NaNO3、酵母膏和温度进行了L9（34）试验设计,以γ-亚麻酸产量为优化目标,直观分析得出影响产量的因素大小顺序为酵母膏、温度、KH2PO4、NaNO3,最终确定最佳的培养条件：酵母膏5 g/L、25℃、KH2PO41 g/L、NaNO33 g/L。在此条件下,检测得出在第5天γ-亚麻酸产量达到0.968 g/L,相比优化前（0.583 g/L）提高了66%。     

Production of medium-chain-length polyhydroxyalkanoates by high-cell-density cultivation of Pseudomonas putida under phosphorus limitation

High-cell-density fed-batch cultures of Pseudomonas putida were carried out for the production of medium-chain-length polyhydroxyalkanoates (PHAs) using oleic acid as a carbon source. By employing an optimal feeding strategy without the limitation of any nutrient, a high cell concentration of 173 g/L was achieved, but the PHA concentration and PHA content were only 32.3 g/L and 18.7 wt%, respectively. To increase the PHA concentration and content, phosphorus limitation was applied during fed-bath culture by reducing the initial KH(2)PO(4) concentration. When the initial KH(2)PO(4) concentration was reduced to 4 g/L, cell concentration, PHA concentration, and PHA content obtained in 38 h were 141 g/L, 72. 6 g/L, and 51.4 wt%, respectively, resulting in a high productivity of 1.91 g PHA/L per hour.     

林可霉素生物合成培养基的优化

以花生粉和棉籽蛋白粉取代了原培养基中的黄豆饼粉,采用响应面法对林可霉素产生菌的发酵培养基进行了优化.首先通过单因素试验及正交实验确定替代氮源及其浓度,采用Plackett-Burman实验分析各因素的主效应,选出对响应值影响较大的3个因素,即花生粉、K2HPO4和玉米浆.对这些因素做爬坡实验,确定三个重要因素的中心点浓...     

Biosynthesis and accumulation of rosmarinic acid in suspension cultures ofColeus blumei

This communication reviews data on the accumulation and biosynthesis of rosmarinic acid in cell suspension cultures ofColeus blumei. The influence of the medium, mainly the carbohydrate source on growth and rosmarinic acid production in these cell cultures is described. The biosynthetic pathway of rosmarinic acid was elucidated inColeus blumei cell cultures: eight enzymatic activities are involved in the transformation of the precursors phenylalanine and tyrosine to the end product rosmarinic acid.Abbreviations CAH cinnamic acid 4-hydroxylase - 4CL 4-coumarate:CoA ligase - HPPR hydroxyphenylpyruvate reductase - 3-H hydroxycinnamoyl-hydroxyphenyllactate 3-hydroxylase - 3-H hydroxycinnamoyl-hydroxyphenyllactate 3-hydroxylase - PAL phenylalanine ammonia-lyase - RAS rosmarinic acid synthase (hydroxycinnamoyl-CoA:hydroxyphenyllactate hydroxycinnamoyl transferase) - TAT tyrosine aminotransferase     

被孢霉菌发酵产生花生四烯酸的研究

研究了温度、培养基初始pH、碳源、氮源对被孢霉(Mortierella sp.)产生花生四烯酸的影响。正交试验结果表明,Mortierella sp.M10最佳培养基组成为(g/L):葡萄糖100,酵母膏10,KNO_3 4.0,KH_2PO_4 2.0,CaCl_2·2H_2O 0.1,MgSO_4·7H_2O 0.5,FeCl_3·6H_2O0.015,ZnSO_4·7H_2O 0.0075,CuSO_4·5H_2O 0.0005。采用最佳培养基及发酵条件,细胞干重和花生四烯酸产量分别为33.51g/L和0.827g/L。同时对摇瓶发酵过程进行了分析。     

Warfarin pharmacogenetics: CYP2C9 and VKORC1 genotypes predict different sensitivity and resistance frequencies in the Ashkenazi and Sephardi Jewish populations

Warfarin is a widely used anticoagulant that has a narrow therapeutic range because of both genetic and environmental factors. CYP2C9( *)2 (p.R144C), CYP2C9( *)3 (p.I359L), and the VKORC1 promoter (g.-1639G-->A) polymorphisms occur frequently in patients who are warfarin "sensitive" and require lower doses, whereas patients with VKORC1 missense mutations are warfarin "resistant" and require higher doses. To compare the CYP2C9 and VKORC1 allele and genotype frequencies among 260 Ashkenazi (AJ) and 80 Sephardi Jewish (SJ) individuals, we genotyped six CYP2C9 and eight VKORC1 alleles by using the Tag-It Mutation Detection Kit and PCR-RFLP assays. The "sensitive"CYP2C9( *)2 and ( *)3 alleles had significantly higher frequencies in SJ than in AJ individuals, 0.194 and 0.144 versus 0.127 and 0.081, respectively (p A, underscoring the importance of screening for p.D36Y prior to initiating warfarin anticoagulation in AJ individuals. Taken together, our findings show that approximately 85% of AJ and approximately 90% of SJ individuals have at least one "sensitive" (CYP2C9( *)2, ( *)3, VKORC1 g.-1639G-->A) or "resistant" (VKORC1 p.D36Y) allele, indicating that each group has different warfarin pharmacogenetics and would benefit from genotype-based dose predictions.     

休哈塔假丝酵母HDYXHT-01利用木糖生产乙醇的发酵工艺优化

采用Plackett-Burman (PB) 方法和中心组合设计 (Ccentral composit design,CCD) 对休哈塔假丝酵母Candida shehataeHDYXHT-01利用木糖发酵生产乙醇的工艺进行优化。PB试验设计与分析结果表明：硫酸铵、磷酸二氢钾、酵母粉和接种量是影响木糖乙醇发酵的4个关键因素,以乙醇产量为响应目标,采用CCD和响应面分析法 (Response surface methodology,RSM),确定了木糖乙醇发酵的最佳工艺为：硫酸铵1.73 g/L、磷酸二氢钾3.56 g/L、酵母粉2.62 g/L和接种量5.66％,其他发酵条件为：木糖80 g/L,MgSO4·7H2O 0.1 g/L,pH 5.0,培养温度30 ℃,装液量100 mL/250 mL,摇床转速140 r/min,发酵时间48 h,在该条件下发酵液中乙醇产量可以达到26.18 g/L,比未优化前提高了1.15倍。