Cadmium-Containing Carbonic Anhydrase CDCA1 in Marine Diatom Thalassiosira weissflogii

The Carbon Concentration Mechanism (CCM) allows phytoplakton species to accumulate the dissolved inorganic carbon (DIC) necessary for an efficient photosynthesis even under carbon dioxide limitation. In this mechanism of primary importance for diatoms, a key role is played by carbonic anhydrase (CA) enzymes which catalyze the reversible hydration of CO₂, thus taking part in the acquisition of inorganic carbon for photosynthesis. A novel CA, named CDCA1, has been recently discovered in the marine diatom Thalassiosira weissflogii. CDCA1 is a cambialistic enzyme since it naturally uses Cd²⁺ as catalytic metal ion, but if necessary can spontaneously exchange Cd²⁺ to Zn²⁺. Here, the biochemical and structural features of CDCA1 enzyme will be presented together with its putative biotechnological applications for the detection of metal ions in seawaters.     

Reproductive performance,survival and development of nauplii and copepodites,sex ratio and adult life expectancy of the harpacticoid copepod,Euterpina acutifrons,fed different microalgal diets

A series of laboratory experiments were conducted on the harpacticoid copepod, Euterpina acutifrons, to assess the influence of 10 different microalgal diets (four monoalgal and six mixed algal diets) on several parameters related to its productivity in culture. The four monoalgal diets were the Tahitian strain of Isochrysis sp. (T‐Iso), Pavlova salina (Pav), Tetraselmis chuii (Tet) and the diatom Chaetoceros muelleri (Chaet), the five binary diets were T‐Iso+Tet, Pav+Tet, T‐Iso+Pav, Tet+Chaet and Pav+Chaet, while the tri‐algal diet was T‐Iso+Tet+Pav. All diets were fed to copepods at 1500 μ gC L^?1 and in the case of binary or trialgal diets, carbon concentration was divided equally between the two or three algae offered. Among monoalgal diets, the diatom Chaet was excellent for E. acutifrons. Out of the 10 diets tested, the binary diet Tet+Chaet, which contained the diatom Chaet, was the best for naupliar production of single pair E. acutifrons (19.5±1.7 nauplii female^?1 day^?1), significantly (P<0.05) higher than all other treatments except for the Chaet treatment (P>0.05). Similarly, in the group naupliar production trial (50 adult E. acutifrons per replicate), Tet+Chaet produced a significantly higher number of nauplii (405.8±18.6 nauplii day^?1) than the other treatments (P<0.05). Tet+Chaet further supported the highest naupliar survival (82.0±2.8%) and copepodite survival (89.0±2.8%), while the mono‐algal diet Chaet produced the second highest naupliar (76.7±2.6%) and copepodite survival (83.5±2.6%). In contrast, Pav produced the lowest overall survival at the naupliar stage (30.0±2.9%), significantly lower than all other treatments (P<0.05). While development from newly hatched nauplii to copepodites was not significantly affected by diets, mean development time from nauplius to adult was significantly different among treatments. Mean development time from hatching (naupliar I stage; NI) to the adult stage was the fastest with Tet+Chaet and Chaet (6.8±0.0 days for both treatments), which was significantly faster than that of Pav, T‐Iso Pav+Tet and T‐Iso+Pav+Te treatment (P<0.05). E. acutifrons sex ratio was significantly affected by diets, and always skewed in favour of males. Feeding on Pav resulted in the lowest proportion of females (23.7±1.2%), significantly lower than for six of the other treatments (P<0.05). Adult females had longer average life expectancy than males for all treatments, and were the longest when fed Tet+Chaet (9.5±0.4 days), which was more than twice as long as the shortest lifespan recorded for the Pav treatment (4.2±0.6 days) (P<0.05). In summary, among 10 diets tested, the binary diet Tet+Chaet appeared to support the highest culture productivity of E. acutifrons while the diatom Chaet also performed well as a monoalgal diet.     

海洋硅藻应对低铁胁迫的研究综述

硅藻在海洋中分布广泛,适应于多种生长环境。本文对海洋硅藻应对低铁胁迫的策略进行综述,以期为研究海洋藻类在低铁环境下的生理代谢状态提供参考。     

南极硅藻GJ01谷胱甘肽合成酶法生产谷胱甘肽

以南极微藻中的南极硅藻GJ01(Berkeleya rutilansGJ01)为材料,采用分光光度法研究谷胱甘肽的前体氨基酸等因子对谷胱甘肽合成酶的影响,并在少量可供应ATP的南极硅藻GJ01细胞存在的情况下,对其游离谷胱甘肽合成酶合成谷胱甘肽的条件进行优化。结果表明,南极硅藻GJ01谷胱甘肽合成酶的最适温度为10～25℃,最适pH值为7.0;Glu、Cys、Gly的最适浓度分别为24 mmol.L-1、12 mmol.L-18、mmol.L-1。此外Met对谷胱甘肽的合成也有影响,而且南极硅藻GJ01的谷胱甘肽合成酶还具有Mg2 依赖性。正交实验表明,南极硅藻GJ01游离谷胱甘肽合成酶,产生谷胱甘肽的最理想条件为,温度30℃、Cys浓度10 mmol.L-1、Mg2 浓度30 mmol.L-1、pH 7.5。该方法用于工业化生产谷胱甘肽是可行的。     

刺参大水体人工育苗研究

剌参大水体育苗,采用降低初耳幼体放养密度,适当减少单胞藻投喂量、加大换水量、通气、增设机动进水系统等措施,培育出稚参1911.5万头,幼参258.36万头,单位水体平均出苗量稚参为3.823万头/m~3,幼参为5167头/m~3。以食用鲜酵母为饵的耳状幼体能正常发育变态,稚参出苗量平均达1.6万头/m~3。稚参的附着、成活,成长与附着基的规格有关。     

鲍育苗用底栖硅藻的研究进展

鲍是珍贵的海产贝类之一，具有很高的营养价值。但近几年，我国台湾、福建、广东以及海南等省发生大规模九孔鲍苗掉板死亡的现象，很多鲍鱼场连续多月多批培苗，但或颗粒无收或每批次收获率极低，不到正常的10％。鲍苗掉板已是南方养鲍业中的一个非常普遍的问题，给我国南方各省鲍鱼养殖业带来极大的负面影响。     

Alanine racemase activity in the microalga <Emphasis Type="Italic">thalassiosira</Emphasis> sp.

ABSTRACT:   In this paper, the authors report the detection of alanine racemase activity in the marine diatom Thalassiosira sp. Since the Thalassiosira sp. was cultured under germ-free conditions, it appeared that D-alanine was not derived from bacteria but was produced through catalysis by algal alanine racemase. The rate of conversion of L-alanine to D-alanine was approximately the same as that for the reverse reaction, and the enzyme catalyzed the equilibration of the D- and L-forms. The crude enzyme preparation obtained from the cells at the stationary phase of the growth cycle had an optimal pH of approximately 9.5. The Lineweaver–Burk analysis showed that the K _m for D- and L-alanine was 16.5 mM and 29.4 mM, respectively. It appears that the enzyme is highly specific for D- or L-alanine because it does not catalyze the racemization of other amino acids. In addition, after gel filtration, the enzyme did not require exogenous pyridoxal 5'-phosphate (PLP) for its activity, however, the effects of several chemicals suggest that the enzyme may be PLP-dependent. The enzyme is more similar to that found in invertebrates when compared with that found in bacteria. This is the first report on the occurrence of alanine racemase activity in the microalga Thalassiosira sp.     

Isolation and Characterization of the Diatom Phaeodactylum Δ5-Elongase Gene for Transgenic LC-PUFA Production in Pichia pastoris

The diatom Phaeodactylum tricornutum can accumulate eicosapentaenoic acid (EPA) up to 30% of the total fatty acids. This species has been targeted for isolating gene encoding desaturases and elongases for long-chain polyunsaturated fatty acid (LC-PUFA) metabolic engineering. Here we first report the cloning and characterization of Δ5-elongase gene in P. tricornutum. A full-length cDNA sequence, designated PhtELO5, was shown to contain a 1110 bp open reading frame encoding a 369 amino acid polypeptide. The putative protein contains seven transmembrane regions and two elongase characteristic motifs of FLHXYHH and MYSYY, the latter being typical for microalgal Δ5-elongases. Phylogenetic analysis indicated that PhtELO5 belongs to the ELO5 group, tightly clustered with the counterpart of Thalassiosira pseudonana. Heterologous expression of PhtELO5 in Pichia pastoris confirmed that it encodes a specific Δ5-elongase capable of elongating arachidonic acid and eicosapentaenoic acid. Co-expression of PhtELO5 and IsFAD4 (a ∆4-desaturase from Isochrysis sphaerica) demonstrated that the high-efficiency biosynthetic pathway of docosahexaenoic acid was assembled in the transgenic yeast. Substrate competition revealed that PhtELO5 exhibited higher activity towards n-3 PUFA than n-6 PUFA. It is hypothesized that Phaeodactylum ELO5 may preferentially participate in biosynthesis of transgenic LC-PUFA via a n-3 pathway in the yeast host.