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1.
Yoshikazu Nishiguchi Fumiyoshi Abe Mitsumasa Okada 《Marine biotechnology (New York, N.Y.)》2011,13(2):137-141
The effects of high hydrostatic pressure on lactate dehydrogenase (LDH) activities from two species of hagfish were examined.
LDH from Eptatretus okinoseanus, a deep-sea species, retained 67% of the original activity even at 100 MPa. LDH activity from Eptatretus burgeri, a shallow-sea species, was completely lost at 50 MPa but recovered to the original value at 0.1 MPa. The tetrameric structure
of LDH-A4 from E. okinoseanus did not change at 50 MPa. In contrast, almost all LDH tetramers from E. burgeri dissociated to dimers and monomers at 50 MPa but reverted to tetramers at 0.1 MPa. These results show that the dissociation
of tetramers caused the inactivation of E. burgeri LDH. The difference depends on the number 6 and 10 amino acids. The mechanism of the slight, gradual inactivation of E. okinoseanus LDH at high pressure differs and is probably due to the metamorphosis of its inner structures. 相似文献
2.
R. A. Patzner 《Helgoland Marine Research》1978,31(1-2):180-190
Photo-reception and sensitivity to light were studied in two Japanese hagfishes,Eptatretus burgeri living in shallow water andParamyxine atami living in water of about 100m depth. Both species responded to general illumination by first moving the tail or head and then by swimming. Local illumination revealed that regions most sensitive to light were the skin of the tail in both species and a line of unpigmented skin running down the back ofE. burgeri. The light sensitivity of the lensless eyes, which are situated below the skin, was very weak in both species.P. atami showed shorter reaction time to light thanE. burgeri. No change in skin colour was induced either by almost complete hypophysectomy or by continuous illumination against a white background. Under-water observations with SCUBA revealed that free movingE. burgeri responded well to illumination uncovered during the night, but the ones buried in mud, with only the heads uncovered did not. 相似文献
3.
Marwen Moussa Vincent Espinasse Jean-Marie Perrier-Cornet Patrick Gervais 《Applied microbiology and biotechnology》2009,85(1):165-174
We investigated the influence of cell hydration on the ability of Saccharomyces cerevisiae CBS 1171 to withstand extreme hydrostatic pressure in order to determine the mechanisms involved in cell resistance. Hydration
conditions were modified in two different ways. We first modulated the chemical potential of water by adding glycerol in cell
suspensions. Another procedure consisted in dehydrating cells aerobically and immersing them in perfluorooctane, an innocuous
hydrophobic liquid used as a pressure-transmitting medium, prior to pressure treatments. This original method made it possible
to transmit isostatic pressure to yeast powders without changing the initial water activity (a
w) level at which cells had been equilibrated. The a
w ranged between 0.11 and 0.99. Pressure treatments were applied at levels of up to 600 MPa for 10 min, 24 h, and 6 days. The
dehydration of cells was found to strongly limit, or even prevent, cell inactivation under pressure. Notably, cells suspended
in a water–glycerol mixture with a
w levels of 0.71 or below were completely protected against all pressure treatments. Moreover, cells dehydrated aerobically
survived for 6 days at 600 MPa even when a
w levels were relatively high (up to 0.94). We highlighted the crucial role of water content in determining cellular damage
under pressure. When water is available in a sufficient amount, high pressure induces membrane permeabilization, causing uncontrolled
mass transfers that could lead to death during a prolonged holding under pressure. Possible mechanisms of membrane permeabilization
are discussed. 相似文献
4.
Evolutionary patterns of sequence divergence were analyzed in genes from the conifer genus Taxus (yew), encoding paclitaxel biosynthetic enzymes taxadiene synthase (TS) and 10-deacetylbaccatin III-10β-O-acetyltransferase
(DBAT). N-terminal fragments of TS, full-length DBAT and internal transcribed spacer (ITS) were amplified from 15 closely related Taxus species and sequenced. Premature stop codons were not found in TS and DBAT sequences. Codon usage bias was not found, suggesting that synonymous mutations are selectively neutral. TS and DBAT gene trees are not consistent with the ITS tree, where species formed monophyletic clades. In fact, for both genes, alleles
were sometimes shared across species and parallel amino acid substitutions were identified. While both TS and DBAT are, overall, under purifying selection, we identified a number of amino acids of TS under positive selection based on inference using maximum likelihood models. Positively selected amino acids in the N-terminal
region of TS suggest that this region might be more important for enzyme function than previously thought. Moreover, we identify
lineages with significantly elevated rates of amino acid substitution using a genetic algorithm. These findings demonstrate
that the pattern of adaptive paclitaxel biosynthetic enzyme evolution can be documented between closely related Taxus species, where species-specific taxane metabolism has evolved recently. 相似文献
5.
Recently, the prenyltransferase SirD was found to be responsible for the O-prenylation of tyrosine in the biosynthesis of sirodesmin PL in Leptosphaeria maculans. In this study, the behavior of SirD towards phenylalanine/tyrosine and tryptophan derivatives was investigated. Product
formation has been observed with 12 of 19 phenylalanine/tyrosine derivatives. It was shown that the alanine structure attached
to the benzene ring and an electron donor, e.g., OH or NH2, at its para-position are essential for the enzyme activity. Modifications were possible both at the side chain and the benzene ring.
Enzyme products from seven phenylalanine/tyrosine derivatives were isolated and characterized by MS and NMR analyses including
HSQC and HMBC and proven to be O- or N-prenylated derivatives at position C4 of the benzene rings. K
M
values of six selected derivatives were found in the range of 0.10–0.68 mM. Catalytic efficiencies (K
cat/K
M
) were determined in the range of 430–1,110 s−1·M−1 with l-tyrosine as the best substrate. In addition, 7 of 14 tested tryptophan analogs were also accepted by SirD and converted to
C7-prenylated derivatives, which was confirmed by comparison with products obtained from enzyme assays using a 7-dimethylallyltryptophan
synthase 7-DMATS from Aspergillus fumigatus. 相似文献
6.
7.
Livingstone JR Yoshida I Tarui Y Hirooka K Yamamoto Y Tsutui N Hirasawa E 《Journal of plant research》2002,115(5):393-400
NAD-dependent aminoaldehyde dehydrogenase (AMADH, EC 1.2.1.-) from Avena shoots was purified by DEAE Sephacel, hydroxyapatite, 5′-AMP Sepharose 4B, Mono Q, and TSK-GEL column chromatographies to
homogeneity by the criterion of native PAGE. SDS–PAGE yielded a single band at a molecular mass of 55 kDa. IEF studies showed
a band with a pI value of 5.3. In contrast to AMADHs from other species, the TSK-GEL chromatography showed that Avena AMADH exists as a monomer in the native state. The purified enzyme catalyzed the oxidations of 3-aminopropionaldehyde (APAL),
4-aminobutyraldehyde (ABAL) N-(3-aminopropyl)-4-aminobutyraldehyde (APBAL), and 4-guanidinobutyraldehyde (GBAL), but not of betaine aldehyde or indoleacetaldehyde.
The K
m values for APAL, ABAL, and GBAL were 1.5×10–6, 2.2×10–6, and 1.3×10–5 M, respectively. Although N-terminal amino acid sequence of Avena AMADH could not be determined due to a modification of the amino residue, the sequence of the fragment of AMADH cleaved by
V8 protease showed greater similarity to the barley BADH than to the pea AMADH.
Electronic Publication 相似文献
8.
On the family Brassicaceae, the causal agent responsible for downy mildew disease was originally regarded as a single species,
Peronospora parasitica (now under Hyaloperonospora), but it was recently reconsidered to consist of many distinct species. In this study, 11 specimens of Peronospora
drabae and P. norvegica parasitic on the genus Draba were investigated morphologically and molecularly. Pronounced differences in conidial sizes (P. drabae: 14–20 × 12.5–15.5 μm; P. norvegica: 20–29 × 15.5–22 μm) and 7.8% sequence distance between their ITS1-5.8S-ITS2 rDNA sequences confirmed their status as distinct
species. Based on ITS phylogeny and morphology (monopodially branching conidiophores, flexuous to sigmoid ultimate branchlets,
hyaline conidia and lobate haustoria), the two species unequivocally belong to the genus Hyaloperonospora and not to Peronospora to which they were previously assigned. Therefore, two new combinations, Hyaloperonospora drabae and H. norvegica, are proposed. The two taxa are illustrated and compared using the type specimen for H. norvegica and authentic specimens for H. drabae, which is lectotypified. 相似文献
9.
The pre-steady states of Pseudomonas species lipase inhibitions by p-nitrophenyl-N-substituted carbamates (1–6) are composed of two steps: (1) formation of the non-covalent enzyme–inhibitor complex (E:I) from the inhibitor and the enzyme
and (2) formation of the tetrahedral enzyme–inhibitor adduct (E–I) from the E:I complex. From a stopped-flow apparatus, the
dissociation constant for the E:I complex, KS, and the rate constant for formation of the tetrahedral E–I adduct from the E:I complex, k2 are obtained from the non-linear least-squares of curve fittings of first-order rate constant (kobs) versus inhibition concentration ([I]) plot against kobs=k2+k2[I]/(KS+[I]). Values of pKS, and log k2 are linearly correlated with the σ* values with the ρ* values of −2.0 and 0.36, respectively. Therefore, the E:I complexes are more positive charges than the inhibitors due to
the ρ* value of −2.0. The tetrahedral E–I adducts on the other hand are more negative charges than the E:I complexes due to the
ρ* value of 0.36. Formation of the E:I complex from the inhibitor and the enzyme are further divided into two steps: (1) the
pre-equilibrium protonation of the inhibitor and (2) formation of the E:I complex from the protonated inhibitor and the enzyme. 相似文献
10.
Zhengfang Zhang Yanming Sheng Keyi Jiang Zhao Wang Yuguo Zheng Qing Zhu 《Biotechnology letters》2010,32(4):513-516
A newly isolated Bacillus megaterium with epoxide hydrolase activity resolved racemic glycidyl (o, m, p)-methylphenyl ethers to give enantiopure epoxides in 84–99% enantiomeric excess and with 21–73 enantiomeric ratios. The (S)-enantiomer was obtained from rac-glycidyl (o or m)-methylphenyl ether while the (R)-epoxides was obtained from glycidyl p-methylphenyl ether. The observations are explained at the level by enzyme-substrate docking studies. 相似文献
11.
12.
Epigenetic modifications of histone play important roles for regulation of cell activity, such as cell division, cell death,
and cell differentiation. A SET domain consisting of about 130 amino acids has lysine methyltransferase activity in the presence
of the cosubstrate S-adenosyl-methionine. More than 60 SET domain-containing proteins have been predicted in various organisms.
One of them, the SMYD family genes which contain a SET domain and a zinc-finger MYND domain are reported to regulate cell
cycle and muscle formation. Here we examined the expression and function of smyd1 and 2 in Xenopus. smyd1 and 2 were expressed in various muscle tissues. While smyd1 expression was observed mainly in cardiac muscle and skeletal muscle, smyd2 expression was done abundantly in skeletal muscle and face region. Moreover, by loss-of-function experiments using antisense
morpholino oligonucleotides, it was suggested that smyd1 and 2 related to muscle cells differentiation. 相似文献
13.
Humulus japonicus in communities of Miscanthus sacchariflorus and Phragmites australis can grow large enough to overtop other species in the Amsa-dong floodplain. Because of strong winds and the weight of Humulus, plants of M. sacchariflorus and P. australis fell in mid-August and were subject to decomposition under its dense shading. To assess the effects of H. japonicus on nutrient cycling in these communities, we collected fresh samples of M. sacchariflorus and P. australis in litterbags and decomposed them under H. japonicus for 9 months, beginning in August. Biomass and organic contents from M. sacchariflorus during this incubation period were 49–51% and 44–48%, whereas those of P. australis were 49–61% and 32–52%, respectively. Their annual k values were 1.61–1.74 and 1.46–3.54, respectively. Initial N concentrations in M. sacchariflorus and P. australis were 13 and 20 mg g−1, while C:N ratios were 31 and 21, respectively. These results indicate that H. japonicus is responsible for the collapse of M. sacchariflorus and P. australis in August and also accelerates their nutrient cycling through rapid decomposition, thereby increasing nutrient circulation
in floodplains. 相似文献
14.
Jian-Xia Zhang Kun-Lin Wu Li-Ning Tian Song-Jun Zeng Jun Duan 《Acta Physiologiae Plantarum》2011,33(2):409-417
15.
Ruiling Liu Meiqin Liu Jie Liu Yuzhen Chen Yiyin Chen Cunfu Lu 《Plant Growth Regulation》2010,60(2):163-168
A late embryogenesis abundant protein gene, AmLEA from Ammopiptanthus mongolicus, was introduced into Escherichia coli using the IMPACT™-TWIN system to analyze the possible function of AmLEA under heat and cold stresses. A fusion protein about 38 kD was expressed in E.coli cells harboring pTWIN-LEA after the induction of IPTG by SDS–PAGE analysis and the accumulation of the fusion protein peaked
3 h after IPTG addition when cultured at 37°C. Compared with control cells, the E. coli cells expressing AmLEA fusion protein showed improved chilling and heat resistence, illuminating the protein may play a protective
role in cells under stress conditions. These results suggested the natively unstructured protein, similar to other members
of LEA proteins, has high capacity for binding water and potential protective function against dehydration or action similar
to the cold shock chaperones. 相似文献
16.
Joong-Chul Lee Jeong-Heon Cha Dennis B. Zerbv George C. Stewart 《Current microbiology》2003,47(2):0146-0152
The divIVB operon of Bacillus subtilis includes the cell shape-associated mre genes, including the membrane-associated proteins MreC and MreD. TnphoA mutagenesis was utilized to analyze a topological model for MreC. MreC has a short cytoplasmic amino terminus, a single membrane-spanning domain, and a large carboxy terminal domain which lies externally to the outer leaflet of the cell membrane. Expression of the B. subtilis MreB protein, or the Mre C and D proteins, results in a morphological conversion of the Escherichia coli host cells from a rod to a roughly spherical cell, morphologically similar to mre-negative mutants of E. coli. Immunolocalization of the MreC protein in B. subtilis revealed that this protein is found at the midcell division site of the bacterial cells, consistent with the postulated role of the Mre proteins in the regulation of septum-specific peptidoglycan synthesis. RID= ID= <E5>Correspondence to: </E5>G.C. Stewart; <E5>email:</E5> stewart@vet.ksu.edu Received: 5 August 2002 / Accepted: 7 October 2002 相似文献
17.
Runli He Zhijian Chang Zujun Yang Zongying Yuan Haixian Zhan Xiaojun Zhang Jianxia Liu 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2009,118(6):1173-1180
Powdery mildew resistance from Thinopyrum intermedium was introgressed into common wheat (Triticum aestivum L.). Genetic analysis of the F1, F2, F3 and BC1 populations from powdery mildew resistant line CH5025 revealed that resistance was controlled by a single dominant allele.
The gene responsible for powdery mildew resistance was mapped by the linkage analysis of a segregating F2 population. The resistance gene was linked to five co-dominant genomic SSR markers (Xcfd233, Xwmc41, Xbarc11, Xgwm539 and Xwmc175) and their most likely order was Xcfd233–Xwmc41–Pm43–Xbarc11–Xgwm539–Xwmc175 at 2.6, 2.3, 4.2, 3.5 and 7.0 cM, respectively. Using the Chinese Spring nullisomic-tetrasomic and ditelosomic lines, the
polymorphic markers and the resistance gene were assigned to chromosome 2DL. As no powdery mildew resistance gene was previously
assigned to chromosome 2DL, this new resistance gene was designated Pm43. Pm43, together with the identified closely linked markers, could be useful in marker-assisted selection for pyramiding powdery
mildew resistance genes.
Runli He and Zhijian Chang contributed equally to this work. 相似文献
18.
Kazuyoshi Ohta Hidenori Tanaka Daisuke Yamakawa Hironori Hamasuna Hirohisa Fujimoto 《Journal of industrial microbiology & biotechnology》2011,38(8):967-973
An extracellular xylanase XynI of glycoside hydrolase family 11 from the dimorphic fungus Aureobasidium pullulans ATCC 20524 possesses an N-terminal extension of 34 amino acids (Ohta et al., J. Biosci. Bioeng. 92:262–270, 2001). The N-terminal
extension includes three sites (Ala-X-Ala-X-Ala-X-Ala) that are potentially cleavable by signal peptidase I of Escherichia
coli. The A. pullulans
xynI signal sequence was fused in frame to the mature protein region of the equivalent xylanase gene xynA from the filamentous fungus Penicillium citrinum. The gene fusion xynI::A was inserted into the plasmid pET-26b(+) to yield pEXP401. An E. coli BL21(DE3) transformant harboring the pEXP401 exhibited xylanase activity (per ml of the culture) of 16.8 U in the fraction
of culture supernatant as well as 4.29 U in the fraction of cell-free extract after 12 h of growth with isopropyl-β-d-thiogalactopyranoside at 30°C. N-terminal amino acid sequence analysis of the secreted recombinant proteins revealed cleavage
at four distinct sites within the N-terminal extension of XynI, two of which conformed to the Ala-X-Ala motif prior to the
cleavage site. The XynA proteins secreted into the culture medium showed high specific activities from 506 to 651 U/mg, which
were twofold higher than that of the native enzyme. 相似文献
19.
Yu YJ Wu SC Chan HH Chen YC Chen ZY Yang MT 《Applied microbiology and biotechnology》2008,81(3):523-532
A novel microbial transglutaminase (TGase) from the cultural filtrate of Streptomyces netropsis BCRC 12429 (Sn) was purified. The specific activity of the purified TGase was 18.2 U/mg protein with an estimated molecular
mass of 38 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis. The TGase gene of S. netropsis was cloned and an open reading frame of 1,242 bp encoding a protein of 413 amino acids was identified. The Sn TGase was synthesized
as a precursor protein with a preproregion of 82 amino acid residues. The deduced amino acid sequence of the mature S. netropsis TGase shares 78.9–89.6% identities with TGases from Streptomyces spp. A high level of soluble Sn TGase with its N-terminal propeptide fused with thioredoxin was expressed in E. coli. A simple and efficient process was applied to convert the purified recombinant protein into an active enzyme and showed
activity equivalent to the authentic mature TGase.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
20.
Mammalian herbivores have the potential to alter the competitive relations of woody species, if consumption is unevenly distributed
between species. At elevations above 3,500 m in the southern Ethiopian highlands, vegetation is dominated by Erica arborea and E. trimera. Both species can potentially grow into short trees, but are burnt on a rotation of 6–10 years, and regenerate by re-sprouting
from belowground lignotubers. The regenerating scrub is heavily browsed by cattle. We set up browsing exclosures at three
burnt sites to quantify the impact of browsing over a 3-year period. When protected from browsing, E. trimera had similar or better height growth than E. arborea, but in browsed vegetation, E. arborea instead grew taller. Browsing was more intense on E. trimera in the first years after fire, indicating a difference in palatability between the species. We checked if browse quality
differed, by analysing shoot contents of acid detergent fibre (ADF), protein, phenolics and tannins. Contrary to expectations,
the preferred E. trimera contained more ADF, less protein and had a higher tannin activity than E. arborea. Although the vegetative growth of E. arborea is favoured relative to E. trimera under high browsing pressure, rapid change in abundance would not be expected, since short-interval fire will repeatedly
eradicate any gains in vegetative growth. However, within the typical fire return interval of less than 10 years, E. trimera barely reach a reproductive state, whereas E. arborea flower profusely. Under the current regime of fire and browsing, this may in the long run be more important than differences
in height growth, leading to a gradual increase in the proportion of E. arborea. 相似文献