RELATIONSHIP BETWEEN CELL CYCLE AND LIGHT‐LIMITED GROWTH RATE IN OCEANIC PROCHLOROCOCCUS (MIT9312) AND SYNECHOCOCCUS (WH8103) (CYANOBACTERIA)1

The relationships between growth rate, cell‐cycle parameters, and cell size were examined in two unicellular cyanobacteria representative of open‐ocean environments: Prochlorococcus (strain MIT9312) and Synechococcus (strain WH8103). Chromosome replication time, C, was constrained to a fairly narrow range of values (～4–6 h) in both species and did not appear to vary with growth rate. In contrast, the pre‐ and post‐DNA replication periods, B and D, respectively, decreased with increasing growth rate from maxima of ～30 and 10–20 h to minima of ～4–6 and 2–3 h, respectively. The combined duration of the chromosome replication and postreplication periods (C+D), a quantity often used in the estimation of Prochlorococcus in situ growth rates, varied ～2.4‐fold over the range of growth rates examined. This finding suggests that assumptions of invariant C+D may adversely influence Prochlorococcus growth rate estimates. In both strains, cell mass was the greatest in slowly growing cells and decreased 2‐ to 3‐fold over the range of growth rates examined here. Estimated cell mass at the start of replication appeared to decrease with increasing growth rate, indicating that the initiation of chromosome replication in Prochlorococcus and Synechococcus is not a simple function of cell biomass, as suggested previously. Taken together, our results reflect a notable degree of similarity between oceanic Synechococcus and Prochlorococcus strains with respect to their growth‐rate‐specific cell‐cycle characteristics.     

Secondary product formation in plant tissue cultures

The formation of secondary products in plant tissue culturesis reviewed. The conditions for the enhanced production of secondaryproducts, which include alkaloids, terpenoids, steroids andphenolics, can be regulated in a number of ways. For example,manipulation of secondary product formation is possible by varyingthe nutrient composition of the growth medium, light, temperatureand pH, and by the use of elicitors, permeabilisation and two-stagesystems. Molecular engineering and the use of biomass and large-scaleculture are described along with future prospects for the commercialproduction of secondary products from cell suspension cultures.     

Ultrahigh resolution drug design. II. Atomic resolution structures of human aldose reductase holoenzyme complexed with Fidarestat and Minalrestat: implications for the binding of cyclic imide inhibitors

The X-ray structures of human aldose reductase holoenzyme in complex with the inhibitors Fidarestat (SNK-860) and Minalrestat (WAY-509) were determined at atomic resolutions of 0.92 A and 1.1 A, respectively. The hydantoin and succinimide moieties of the inhibitors interacted with the conserved anion-binding site located between the nicotinamide ring of the coenzyme and active site residues Tyr48, His110, and Trp111. Minalrestat's hydrophobic isoquinoline ring was bound in an adjacent pocket lined by residues Trp20, Phe122, and Trp219, with the bromo-fluorobenzyl group inside the "specificity" pocket. The interactions between Minalrestat's bromo-fluorobenzyl group and the enzyme include the stacking against the side-chain of Trp111 as well as hydrogen bonding distances with residues Leu300 and Thr113. The carbamoyl group in Fidarestat formed a hydrogen bond with the main-chain nitrogen atom of Leu300. The atomic resolution refinement allowed the positioning of hydrogen atoms and accurate determination of bond lengths of the inhibitors, coenzyme NADP+ and active-site residue His110. The 1'-position nitrogen atom in the hydantoin and succinimide moieties of Fidarestat and Minalrestat, respectively, form a hydrogen bond with the Nepsilon2 atom of His 110. For Fidarestat, the electron density indicated two possible positions for the H-atom in this bond. Furthermore, both native and anomalous difference maps indicated the replacement of a water molecule linked to His110 by a Cl-ion. These observations suggest a mechanism in which Fidarestat is bound protonated and becomes negatively charged by donating the proton to His110, which may have important implications on drug design.     

禁牧条件下不同类型草地群落结构特征

利用幂乘方法则模型探讨了鄂尔多斯3种不同类型草地在禁牧情况下的群落结构特征,包括物种组成、物种多样性、生物量和空间分布规律。结果表明:幂乘方法则在解析鄂尔多斯不同类型草地的空间异质性时具有很好的吻合性;羊草(Leymuschinensis)草地、芨芨草(Achnatherum splendens)草地和油蒿(Artemisia ordosica)灌丛草地均比随机分布呈现了较强的空间异质性,群落整体的空间异质性指数表现为油蒿灌丛草地>羊草草地>芨芨草草地;群落整体的物种多样性指数为油蒿灌丛草地>羊草草地>芨芨草草地;L-样方(50cm×50cm)内的平均物种数和物种多样性指数均表现为羊草草地极显著地高于油蒿灌丛草地,油蒿灌丛草地又极显著地高于芨芨草草地(P<0.001);L-样方内的平均生物量表现为油蒿灌丛草地极显著地高于芨芨草草地(P<0.001),而羊草草地与油蒿灌丛草地以及芨芨草草地之间没有显著性差异;这3种类型的草地其L-样方内的平均生物量和物种多样性指数都随着群落整体空间异质性指数的增大而增大。     

Organic matter and nitrogen accumulation and nitrogen fixation during early ecosystem development in Hawaii

We used a chronosequence comprised of 10 y, 52 y and 142 yold `a'a lava flows on Mauna Loa, Hawaii, to determine theaccumulation of organic matter and nitrogen and rates of nitrogenfixation through time. The mass of organic matter (live and deadbiomass and soil organic matter) on the 1984, 1942 and 1852 lavaflows was 0.6, 2.2 and 7.6 kg m^{– 2}, respectively, while total N was 4.8, 10.9 and 85.7 g m^{– 2}.We estimated the total rates of nitrogen fixation for thethree different aged ecosystems using an acetylene reduction assaycalibrated with ¹⁵N incubations. While mean rates of total N fixation remained largely constant across the three sites – between2.0 and 3.1 kg ha^{– 1} y^{– 1} – the most important sources of N fixation changed. On the 10 y flow, the most important fixer was the pioneering cyanolichen, Stereocaulon vulcani. After 52 years ofecosystem development, the most important N fixer was a cyanoalga,while after 142 years, the predominant N fixers were heterotrophicbacteria associated with leaf litter, twigs and detritus. The totalamount of N accumulated after 52 years of ecosystem development wasequivalent to cumulative inputs through biological N fixation. After 142 years, however, cumulative inputs from N fixation couldonly account for between 27–59% of the total nitrogen accrued.We used fertilizer additions of all essential nutrients otherthan N to test whether the availability of lithophilic nutrientsregulated rates of N fixation in early ecosystem development. Ratesof nitrogen fixation by the lichen, S. vulcani, approximately doubled when fertilized on the 1984 and 1942 flows. Rates of N-fixation by heterotrophic nitrogen fixing bacteria on leaf litter ofMetrosideros polymorpha also increased significantly when fertilized with lithophilic nutrients. These findings suggest that weathering rates of lava in part regulate rates of nitrogen fixation in these young ecosystems.     

Structure of Novel Enzyme in Mannan Biodegradation Process 4-O-β-d-Mannosyl-d-Glucose Phosphorylase MGP

The crystal structure of a novel component of the mannan biodegradation system, 4-O-β-d-mannosyl-d-glucose phosphorylase (MGP), was determined to a 1.68-Å resolution. The structure of the enzyme revealed a unique homohexameric structure, which was formed by using two helices attached to the N-terminus and C-terminus as a tab for sticking between subunits. The structures of MGP complexes with genuine substrates, 4-O-β-d-mannosyl-d-glucose and phosphate, and the product d-mannose-1-phosphate were also determined. The complex structures revealed that the invariant residue Asp131, which is supposed to be the general acid/base, did not exist close to the glycosidic Glc-O4 atom, which should be protonated in the catalytic reaction. Also, no solvent molecule that might mediate a proton transfer from Asp131 was observed in the substrate complex structure, suggesting that the catalytic mechanism of MGP is different from those of known disaccharide phosphorylases.     

LacZ β‐galactosidase: Structure and function of an enzyme of historical and molecular biological importance

This review provides an overview of the structure, function, and catalytic mechanism of lacZ β‐galactosidase. The protein played a central role in Jacob and Monod's development of the operon model for the regulation of gene expression. Determination of the crystal structure made it possible to understand why deletion of certain residues toward the amino‐terminus not only caused the full enzyme tetramer to dissociate into dimers but also abolished activity. It was also possible to rationalize α‐complementation, in which addition to the inactive dimers of peptides containing the “missing” N‐terminal residues restored catalytic activity. The enzyme is well known to signal its presence by hydrolyzing X‐gal to produce a blue product. That this reaction takes place in crystals of the protein confirms that the X‐ray structure represents an active conformation. Individual tetramers of β‐galactosidase have been measured to catalyze 38,500 ± 900 reactions per minute. Extensive kinetic, biochemical, mutagenic, and crystallographic analyses have made it possible to develop a presumed mechanism of action. Substrate initially binds near the top of the active site but then moves deeper for reaction. The first catalytic step (called galactosylation) is a nucleophilic displacement by Glu537 to form a covalent bond with galactose. This is initiated by proton donation by Glu461. The second displacement (degalactosylation) by water or an acceptor is initiated by proton abstraction by Glu461. Both of these displacements occur via planar oxocarbenium ion‐like transition states. The acceptor reaction with glucose is important for the formation of allolactose, the natural inducer of the lac operon.