多酚氧化酶酶学特性研究及其应用进展

多酚氧化酶广泛存在于各类果蔬植物中,其活性调控对于果蔬加工、保藏及茶叶初加工过程具有重要的作用。着重归纳了多酚氧化酶的催化活性及其影响因素,结合果蔬抑褐保鲜及茶叶加工的应用实例进行了分类综述,并指出了多酚氧化酶在未来生物技术中的价值空间。     

The two actin-interacting protein 1 genes have overlapping and essential function for embryonic development in Caenorhabditis elegans

Disassembly of actin filaments by actin-depolymerizing factor (ADF)/cofilin and actin-interacting protein 1 (AIP1) is a conserved mechanism to promote reorganization of the actin cytoskeleton. We previously reported that unc-78, an AIP1 gene in the nematode Caenorhabditis elegans, is required for organized assembly of sarcomeric actin filaments in the body wall muscle. unc-78 functions in larval and adult muscle, and an unc-78-null mutant is homozygous viable and shows only weak phenotypes in embryos. Here we report that a second AIP1 gene, aipl-1 (AIP1-like gene-1), has overlapping function with unc-78, and that depletion of the two AIP1 isoforms causes embryonic lethality. A single aipl-1-null mutation did not cause a detectable phenotype. However, depletion of both unc-78 and aipl-1 arrested development at late embryonic stages due to severe disorganization of sarcomeric actin filaments in body wall muscle. In vitro, both AIPL-1 and UNC-78 preferentially cooperated with UNC-60B, a muscle-specific ADF/cofilin isoform, in actin filament disassembly but not with UNC-60A, a nonmuscle ADF/cofilin. AIPL-1 is expressed in embryonic muscle, and forced expression of AIPL-1 in adult muscle compensated for the function of UNC-78. Thus our results suggest that enhancement of actin filament disassembly by ADF/cofilin and AIP1 proteins is critical for embryogenesis.     

A flow cytometry-based screening system for directed evolution of proteases

Proteases are industrially important enzymes but often have to be improved for their catalytic efficiency and stabilities to suit applications. Flow cytometry screening technology based on in vitro compartmentalization in double emulsion had been developed and applied on directed evolution of paraoxonase and β-galactosidase. Further advancements of flow cytometry-based screening technologies will enable an ultra-high throughput of variants offering novel opportunities in directed enzyme evolution under high mutational loads. For the industrially important enzyme class of proteases, a first flow cytometry-based screening system for directed protease evolution has been developed based on an extracellular protease-deficient Bacillus subtilis strain (WB800N), a model protease (subtilisin Carlsberg), and a water-in-oil-in-water double-emulsion technology. B. subtilis WB800N cells are encapsulated in double emulsion with a fluorogenic substrate (rhodamine 110-containing peptide), allowing the screening of protease variants in femtoliter compartments at high throughput. The protease screening technology was validated by employing an epPCR mutant library with a high mutational load and screened for increased resistance toward the inhibitor antipain dihydrochloride. A variant (K127R, T237P, M239I, I269V, Y310F, I372V) with an improved relative resistance was isolated from a small population of active variants, validating the reported protease flow cytometry screening technology for increased inhibitor resistance.     

Comparison of mRNA expression patterns of class B scavenger receptors in BV2 microglia upon exposure to amyloidogenic fragments of beta-amyloid and prion proteins

The inflammatory responses in Alzheimer's disease and prion diseases are dominated by microglia activation. Scavenger receptors have been recently related to the innate immune activation of microglia initiated by endogenous ligands. In this study, we investigated mRNA expression patterns of B class scavenger receptors CD36 and scavenger receptor B1 (SR-B1) in BV2 microglia upon exposure to amyloid fibril Aβ(1-42) and PrP(106-126), respectively. CD36 and SR-B1 showed similar mRNA expression patterns following each treatment. PrP(106-126) induced a rapid increase of CD36 and SR-B1 mRNA levels in the treated microglia, whereas Aβ(1-42) induced a delayed but persistent increase in the mRNA expression of CD36 and SR-B1. These results suggest a possible involvement of CD36 and SR-B1 in microglial interaction with amyloidogenic fragments of beta-amyloid and prion proteins.     

Identification of two major QTL for yellow seed color in two crosses of resynthesized <Emphasis Type="Italic">Brassica napus</Emphasis> line No. 2127-17

Yellow seed color, which results from a thinner seed coat, is associated with improved feed quality of rapeseed (Brassica napus L.) meal and increased oil and protein content. As this trait follows various genetic models under different genetic backgrounds, a study was performed in two genetic backgrounds to gain a better understanding of the genetic mechanisms underlying yellow seed color. The quantitative trait locus (QTL) analysis was undertaken using two crosses, Quantum ×No. 2127-17 (HZ-1) and No. 2127-17 × 94,570 (HZ-2). In the HZ-1 population, three putative QTL were detected in linkage groups N18, N5, and N3, respectively. For all of them, yellow seed color arose from the No. 2127-17 alleles. Of these QTL, the one in linkage group N18 (Bnsc-18a) explained more than half of the phenotypic variation. In the HZ-2 population, three QTL were found in linkage groups N9, N18, and N8, respectively. Of these QTL, that in linkage group N9 (Bnsc-9a) explained more than half of the phenotypic variation, whereas the QTL Bnsc-18a had a low seed color value and explained only 9.03–11.72% of the phenotypic variation. Bulked segregant analysis (BSA) of the extremes of a BC1 population derived from the cross of No. 2127-17 × 94,570 (HZ-3) identified one major gene that was identical with the QTL Bnsc-9a detected in the HZ-2 population. The QTL Bnsc-18a was common in the HZ-1 and HZ-2 populations, and the others were population-specific. These results suggested that different black-seeded forms had different seed color genes.     

Comparative genomics of the social amoebae <Emphasis Type="Italic">Dictyostelium discoideum</Emphasis> and <Emphasis Type="Italic">Dictyostelium purpureum</Emphasis>

Background

The social amoebae (Dictyostelia) are a diverse group of Amoebozoa that achieve multicellularity by aggregation and undergo morphogenesis into fruiting bodies with terminally differentiated spores and stalk cells. There are four groups of dictyostelids, with the most derived being a group that contains the model species Dictyostelium discoideum.

Results

We have produced a draft genome sequence of another group dictyostelid, Dictyostelium purpureum, and compare it to the D. discoideum genome. The assembly (8.41 × coverage) comprises 799 scaffolds totaling 33.0 Mb, comparable to the D. discoideum genome size. Sequence comparisons suggest that these two dictyostelids shared a common ancestor approximately 400 million years ago. In spite of this divergence, most orthologs reside in small clusters of conserved synteny. Comparative analyses revealed a core set of orthologous genes that illuminate dictyostelid physiology, as well as differences in gene family content. Interesting patterns of gene conservation and divergence are also evident, suggesting function differences; some protein families, such as the histidine kinases, have undergone little functional change, whereas others, such as the polyketide synthases, have undergone extensive diversification. The abundant amino acid homopolymers encoded in both genomes are generally not found in homologous positions within proteins, so they are unlikely to derive from ancestral DNA triplet repeats. Genes involved in the social stage evolved more rapidly than others, consistent with either relaxed selection or accelerated evolution due to social conflict.

Conclusions

The findings from this new genome sequence and comparative analysis shed light on the biology and evolution of the Dictyostelia.

     

Supramolecular copolymer micelles based on the complementary multiple hydrogen bonds of nucleobases for drug delivery

Novel supramolecular copolymer micelles with stimuli-responsive abilities were successfully prepared through the complementary multiple hydrogen bonds of nucleobases and then applied for rapid intracellular release of drugs. First, both adenine-terminated poly(ε-caprolactone) (PCL-A) and uracil-terminated poly(ethylene glycol) (PEG-U) were synthesized. The supramolecular amphiphilic block copolymers (PCL-A:U-PEG) were formed based on multiple hydrogen bonding interactions between PCL-A and PEG-U. The micelles self-assembled from PCL-A:U-PEG were sufficiently stable in water but prone to fast aggregation in acidic condition due to the dynamic and sensitive nature of noncovalent interactions. The low cytotoxicity of supramolecular copolymer micelles was confirmed by MTT assay against NIH/3T3 normal cells. As a hydrophobic anticancer model drug, doxorubicin (DOX) was encapsulated into these supramolecular copolymer micelles. In vitro release studies demonstrated that the release of DOX from micelles was significantly faster at mildly acid pH of 5.0 compared to physiological pH. MTT assay against HeLa cancer cells showed DOX-loaded micelles had high anticancer efficacy. Hence, these supramolecular copolymer micelles based on the complementary multiple hydrogen bonds of nucleobases are very promising candidates for rapid controlled release of drugs.     

退化红壤丘陵区森林凋落物初始化学组成与分解速率的关系

通过小盆+凋落袋控制试验,研究了我国南方退化红壤丘陵区8种森林凋落物和4种混合凋落物初始化学组成与分解速率的关系.结果表明:阔叶凋落物中的氮、磷、钾、镁含量显著高于针叶凋落物,木质素、碳含量显著低于针叶凋落物;凋落物分解速率与凋落物初始氮、磷、钾、镁含量呈显著正相关,与凋落物初始碳、木质素含量以及木质素/氮、木质素/磷和碳/磷值呈显著负相关;木质素含量解释了凋落物分解速率变异的54.3%,是影响分解速率的最关键因子,凋落物碳、氮、磷含量也与分解速率密切相关,它们与木质素含量一起可解释分解速率变异的81.4%.在退化红壤丘陵区植被恢复过程中,低木质素含量、高氮磷含量的阔叶物种的引入有利于加速凋落物的分解速率和土壤肥力的恢复进程.     

长期定位施肥对石灰性紫色水稻土古菌群落结构的影响

为了认识长期施肥对石灰性紫色水稻土培肥和肥力演化的作用,结合变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)和限制性酶切片段长度多态性(RFLP)技术,研究了稻麦轮作下农家肥(M)、氮肥+农家肥(NM)、氮磷肥+农家肥(NPM)、氮磷钾肥+农家肥(NPKM)、无肥(CK)、氮肥(N)、氮磷肥(NP)、氮磷钾肥(NPK)等不同施肥制度对石灰性紫色水稻土古菌群落结构的影响.研究结果表明,长期定位施肥明显影响土壤中的古菌组成.在长期施用氮肥+农家肥、氮磷肥和氮磷钾肥+农家肥处理的土壤中,古菌多样性指数低于农家肥、氮磷肥+农家肥、无肥、氮肥和氮磷钾肥处理.在DGGE图谱的基础上,分别选择种植水稻和小麦的氮磷钾肥处理土壤样品,对古菌克隆子的16S rDNA序列进行了系统发育分析,发现水稻七古菌与各种土壤及水体环境的古菌极其相似.对DGGE图谱的聚类分析发现,不管是种植水稻还是小麦,8种施肥处理的古菌都聚在3个群里.种植水稻时,M和NPK处理下的土壤古菌聚成第一个群,NP处理下的聚成第二个群,另外5种施肥处理(NPKM,NM,Ck,N和NPM)聚成第三个群.种植小麦时,NPKM和M处理下的土壤古菌聚成一个群,NP处理下的聚成第二个群,N、NPK、NM、NPM和CK处理下的聚成第三个群.聚类分析结果显示,作物类型会影响土壤古菌群落结构.