Isolation of an Unusually Stable Cis-Isomer of Alloxanthin from a Bleached Autolysed Culture of Chroomonas salina Grown Photoheterotrophically on Glycerol. Observations on Cis-Trans Isomerization of Alloxanthin

SYNOPSIS. A new carotenoid, manixanthin , was isolated from a bleached, autolysed culture of the marine cryptomonad Chroomonas salina grown on glycerol in light, after the culture showed disappearance of the chlorophylls and carotenoids normally produced. Manixanthin lacked carbonyl, ester, and epoxy groups in chemical tests, whilst its spectroscopic properties indicated a conjugated nonaene chromophore. In its chromatographic mobility, manixanthin closely resembled alloxanthin, the major normal xanthophyll of the cryptomonad. The iodine-catalysed photostereomutation of manixanthin produced the same equilibrium mixture of cis -isomers as that given by similar treatment of alloxanthin. This mixture of isomers completely lacked the all-trans alloxanthin and contained ∼60% manixanthin. It was concluded that manixanthin is an unusually stable cis -isomer of alloxanthin, and is presumably produced in the culture medium by the action of light on alloxanthin released by autolysed cells of the cryptomonad. The unusual stability of manixanthin relative to alloxanthin was investigated by effecting partial stereoisomerizations under conditions using either of heat, light, or iodine, and evidence was obtained for a series of at least 10 isomers with a trend of stability increase from alloxanthin to manixanthin. It was inferred that manixanthin may be 9,9'-di cis -alloxanthin.     

Wax Ester Production by the Marine Cryptomonad Chroomonas salina Grown Photoheterotrophically on Glycerol

SYNOPSIS. An age-autolyzed culture of Chroomonas salina , grown under cool-white light with glycerol, produced waxy lipid constituting about 44% of total matter harvested. This lipid was composed of 87% wax ester, 9% triglyceride, 3% polar lipid and 1% hydrocarbon. The major wax ester species were identified by total carbon number as C₂₆(28%), C₂₈(35%), C₃₀(15%). The main fatty acid components of the wax esters were 12:0 (39%), 14:0 (30%), 16:0 (14%), while the main alcohols were 14:0 (53%) and 16:0 (40%). The hydrocarbon fraction showed saturated paraffins ranging from C₁₇ to C₃₃, with odd-numbered chain components predominating. No polyunsaturated components were detected in the wax ester or hydrocarbon fractions. This is the first record of wax ester production by a cryptomonad or a marine phytoplankter.     

Effect of Ca channel blockers on glycerol levels in Dunaliella tertiolecta under hypoosmotic stress

The role of Ca²⁺ in glycerol dissimilation under hypoosmotic stress in the halotolerant alga Dunaliella tertiolecta was investigated using a pharmacological approach. A stretch-activated Ca²⁺ channel blocker, GdCl₃, inhibited glycerol dissimilation under hypoosmotic stress. However, addition of voltage-dependent Ca²⁺ channel blockers and inhibitors of mitochondrial and endoplasmic reticulum Ca²⁺ channels did not affect the glycerol dissimilation under hypoosmotic stress. The results of the present study suggest that the influx of Ca²⁺ from the extracellular space via the stretch-activated Ca²⁺ channels localized in the plasma membrane is required for the transduction of osmotic signal of D. tertiolecta.     

Effects of High K+ and Alkaline pH on Ultrastructure of Dunaliella salina Chloroplasts

It was reported that the growth of Dunaliella salina Teod. cultured in medium containing 1 mol/L NaC1 was almost completely inhibited by the addition of 100 mmol/L KC1. The high K+ (100 mmol/L KC1) treatment also significantly inhibited the photosynthetic rate of D. salina and decreased chlorophyll contents in algae. This study focuses on possible effects of high K+ or alkaline pH on the ultrastructural change of chloroplasts in D. salina. After D. salina was cultured in a medium containing 100 n,anol/L KC1 or in a medium with alkaline pH for 8 to 10 days, dramatic ultrastructural changes occurred in the chloroplasts including thylakoid swelling, volume increase of chloroplast, and significant accumulation of starch grains in chloroplasts. The results are consistent with our previous report indicating that the ultrastmctuml changes in chloroplast under high K + or alkaline pH may lead to an inhibitory effects on photosynthesis and overall growth of D. salina.     

盐生杜氏藻甘油-3-磷酸脱氢酶的分离纯化及其特性的研究

利用ＰＥＧ分级,ＤＥＡＥ离子交换层析,ＢｌｕｅＳｅｐｈａｒｏｓｅ拟亲和层析,ＭｏｎｏＱ离子交换层析等手段,分离纯化盐生杜氏藻（Ｄｕｎａｌｉｅｌａｓａｌｉｎａ（Ｄｕｎａｌ）Ｔｅｏｄ．）甘油三磷酸（Ｇ３Ｐ）脱氢酶（ＥＣ１．１．１．８）,得到比活为１２．６Ｕ／ｍｇ的电泳纯的酶,并对此酶的生化特性进行了研究。４％～２０％非变性聚丙烯酰胺梯度凝胶电泳测得全酶分子量约为２７０ｋＤ,ＳＤＳＰＡＧＥ表明该酶只有一种分子量约为６５ｋＤ的亚基,据此推测该酶应为同四聚体。酶催化磷酸二羟丙酮（ＤＨＡＰ）还原的最适ｐＨ值为７．５,催化Ｇ３Ｐ脱氢的最适ｐＨ值为１０。该酶对４个底物还原型辅酶Ⅰ（ＮＡＤＨ）,二磷酸吡啶核苷酸（ＤＨＡＰ）,辅酶Ⅰ（ＮＡＤ）,Ｇ３Ｐ的表观Ｋｍ值分别为６３μｍｏｌ／Ｌ,２７２μｍｏｌ／Ｌ,１．５３ｍｍｏｌ／Ｌ,６．５２ｍｍｏｌ／Ｌ。该酶在保存过程中易失活。ＮＡＤＨ能降低酶失活的速度,而ＮＡＤ则不然。低浓度ＮａＣｌ对酶略有保护作用,但高浓度ＮａＣｌ加快酶的失活,且浓度越高效应越明显。     

葡萄糖对杜氏盐藻3-磷酸甘油脱氢酶活性的影响

杜氏盐藻是一种抗渗透能力强的单细胞绿藻,甘油在其渗透调节过程中具有重要作用。葡萄糖对杜氏盐藻细胞数量的增加效果不明显,但对盐藻细胞内甘油积累有显著促进作用,在0～15g/L范围内葡萄糖的浓度与胞内甘油积累显著相关(R2=0.9604,P=0.01);葡萄糖浓度达到15g/L时,胞内甘油积累量达到最高值7.80pg/cell,是对照的1.88倍,胞内甘油积累量与葡萄糖的消耗量极显著相关(R2=0.9982,P=0.01)。葡萄糖对盐藻细胞内总蛋白、3-磷酸甘油脱氢酶(GPDH)酶活和比活都有显著影响,在15g/L葡萄糖时这3个值达到最大值,分别是对照的1.354、4.384、3.229倍。数据显示葡萄糖浓度在15g/L时细胞内蛋白质含量增加不多,但GPDH酶活和比活却大幅度增加;葡萄糖导致的渗透压的变化可能诱导新的同功酶的合成。     

葡萄糖诱导绿色杜氏藻转录组及相关通路差异分析

绿色杜氏藻是研究耐盐机理的模式绿藻．葡萄糖不仅是营养物质,而且还是信号物质．目前,对绿色杜氏藻转录组、糖处理后差异表达基因和β-胡萝卜素生物合成途径关键基因表达还不清楚．本研究通过Illumina HiSeqTM 2000高通量测序,获得葡萄糖处理和未处理绿色杜氏藻转录组信息．利用P value值和差异倍数对样本进行差异表达分析,共111条转录本存在差异表达,3条为上调转录本,108条为下调转录本．利用RT-qPCR检验差异表达分析的准确性. 结果表明,转录本表达结果与转录组分析结果一致．GO功能富集结果表明,71条下调转录本与代谢相关,占所有下调转录本的65.74%．KEGG富集分析结果表明,21条KEGG通路含89条下调转录本,14条通路与代谢相关．代谢中通路最多的为能量代谢（6条）,含63条下调转录本．能量代谢中与光合作用相关的下调转录本最多,为29条．通过分析找到2条与β-胡萝卜素生物合成相关通路（MVA/MEP途径及β-胡萝卜素合成途径）,并发现通路的关键基因hmgs、dxs、dxr、psy、pds、chyb,对其进行差异表达分析,均不存在差异表达．研究表明,葡萄糖抑制了绿色杜氏藻光合作用,代谢受阻,但未影响β-胡萝卜素生物合成相关通路及关键基因．     

Assessment of carotenoid production by Dunaliella salina in different culture systems and operation regimes

The effect of operation regime and culture system on carotenoid productivity by the halotolerant alga Dunaliella salina has been analyzed. Operation strategies tested included batch and semi continuous regime, as well as a two-stage approach run simultaneously in both, open tanks and closed reactor. The best results were obtained with the closed tubular photobioreactor. The highest carotenoid production (328.8 mg carotenoid l⁻¹ culture per month) was achieved with this culture system operated following the two-stage strategy. Also, closed tubular photobioreactor provided the highest carotenoid contents (10% of dry weight) in Dunaliella biomass and β-carotene abundance (90% of total carotenoids) as well as the highest 9-cis to all-trans β-carotene isomer ratio (1.5 at sunrise).