酶法水解中国毛虾的研究(英文)

比较了4种常用蛋白酶对中国毛虾的酶解效果,筛选出胰蛋白酶和枯草杆菌蛋白酶为最佳用酶,并利用正交试验优化了水解条件,在各自最佳水解条件下采用先加胰蛋白酶再加枯草杆菌蛋白酶的酶解方式对中国毛虾进行联合双酶水解,水解度可达67 .83%。     

Enzymatic hydrolysis of defatted mackerel protein with low bitter taste

Ultrasound-assisted solvent extraction was confirmed as a novel,effective method for separating lipid from mackerel pro-tein,resulting in a degreasing rate (DR) of 95% and a nitrogen recovery (NR) of 88.6%.To obtain protein hydrolysates with high ni-trogen recovery and low bitter taste,enzymatic hydrolysis was performed using eight commercially available proteases.It turned out that the optimum enzyme was the ‘Mixed enzymes for animal proteolysis’.An enzyme dosage of 4%,a temperature of 50℃,and a hydrolysis time of 300 min were found to be the optimum conditions to obtain high NR (84.28%) and degree of hydrolysis (DH,16.18%) by orthogonal experiments.Glutamic acid was the most abundant amino acid of MDP (defatted mackerel protein) and MDPH (defatted mackerel protein hydrolysates).Compared with the FAO/WHO reference protein,the essential amino acid chemical scores (CS) were greater than 1.0 (1.0 1.7) in MDPH,which is reflective of high nutritional value.This,coupled with the light color and slight fishy odor,indicates that MDPH would potentially have a wide range of applications such as nutritional additives,functional ingredients,and so on.     

Properties of Klebsiella phage P13 and associated exopolysaccharide depolymerase

The bacteriophage P13 that infects Klebsiella serotype K13 contains a heat-stable depolymerase capable of effective degradation of exopolysaccharide(EPS) produced by this microorganism. In this study, the titer of phage P13, initially 2.0 × 107 pfu mL-1, was found increasing 20 min after infection and reached 5.0 × 109 pfu mL-1 in 60 min. Accordingly, the enzyme activity of depolymerase approached the maximum 60 min after infection. Treatment at 70℃ for 30 min inactivated all the phage, but retained over 90% of the depolymerase activity. Addition of acetone into the crude phage lysate led to precipitation of the protein, with a marked increase in bacterial EPS degradation activity and a rapid drop in the titer of phage. After partial purification by acetone precipitation and ultrafiltration centrifugation, the enzyme was separated from the phage particles, showing two components with enzyme activity on Q-Sepharose Fast Flow. The soluble enzyme had an optimum degradation activity at 60℃ and pH 6.5. Transmission electron microscopy demonstrated that the phage P13 particles were spherical with a diameter of 50 nm and a short stumpy tail. It was a double-strand DNA virus consisting of a nucleic acid molecule of 45976 bp. This work provides an efficient purification operation including thermal treatment and ultrafiltration centrifugation, to dissociate depolymerase from phage particles. The characterization of phage P13 and associated EPS depolymerase is beneficial for further application of this enzyme.     

DETERMINATION OF AMINO ACIDS IN TWO POLYSIPHONIA SPECIES AND STUDY OF ENZYMATIC HYDROLYSIS METHOD

The total content ot the nch amino acids in two common red algae, Potystpnoma urceolata and Polysiphonia japonica growing in the Qingdao seashore were determined. The algae powder was hydrolyzed by 6 mol/L HC1 at 110℃ for 48 h and determined by amino acid analyzer. The content was 25.35% and 24.16% , respectively, much higher than that of some other species. In addition, a nutritive liquid with abundant amino acids was prepared (by the enzymatic hydrolysis method using Polysiphonia urceolata ) as raw material for a kind of health beverage. The dried seaweed was decolored by 0.25% KMnO4 and 0.5% active carbon, then enzymalized. In the selection of enzymalizing condition, the orthogonal experimental design was used. Four factors including kinds of enzyme, quantity of enzyme, temperature and time were studied at 3 levels. According to the orthogonal design results, we can choose an optimal condition: hydrolyzing at 45℃ by neutral proteinase (0.25% , w/w) for 2 h, adjusting pH to 8.5, then adding trypsin (0.25% , w/w) and hydrolyzing for 2 h. Finally the above solution was alkalized by NaOH and neutralized by casein. After the hydrolyzed liquid was filtered and concentrated, suitable additives were added. The final products contain rich amino acids.     

Purification and characterization of iron-cofactored superoxide dismutase from Enteromorpha linza

A superoxide dismutase was purified from Enteromorpha linza using a simple and safe procedure, which comprised phosphate buffer extraction, ammonium sulphate precipitation, ion exchange chromatography on Q-sepharose column, and gel filtration chromatography on Superdex 200 10/300GL. The E. linza superoxide dismutase （E/SOD） was purified 103.6-fold, and a yield of 19.1% and a specific activity of 1 750 U/rag protein were obtained. The SDS-PAGE exhibited E/SOD a single band near 23 kDa and the gel filtration study showed E/SOD＇s molecular weight is near 46 kDa in nondenatured condition, indicating it＇s a homodimeric protein. E/SOD is an iron-cofactored superoxide dismutase （Fe-SOD） because it was inhibited by hydrogen peroxide, insensitive to potassium cyanide. The optimal temperature for its maximal enzyme activity was 35℃, and it still had 29.8% relative activity at 0℃, then E/SOD can be classified as a cold-adapted enzyme. E/SOD was stable when temperature was below 40℃ or the pH was within the range of 5 10. The first 11 N-terminal amino acids orE/SOD were ALELKAPPYEL, comparison of its N-terminal sequence with other Fe-SOD N-terminal sequences at the same position suggests it is possibly a chloroplastic Fe-SOD.     

酶解蓝圆鲹鱼肉蛋白制备小分子肽的工艺研究

以蓝圆鲹为原料,酶解蓝圆鲹鱼肉蛋白制备小分子肽。运用响应面试验设计优化蓝圆鲹蛋白酶解的工艺条件,探讨了酶种类和添加量、液固比、酶解温度、酶解时间对多肽提取率的影响。结果表明:选用木瓜蛋白酶,添加量为0.10%,液固比2∶1,酶解温度54.4℃,酶解时间3.18 h下,多肽提取率为32.35%。     

Effects of different enzymatic hydrolysis methods on the bioactivity of peptidoglycan in Litopenaeus vannamei

The effects of different hydrolysis methods on peptidoglycan(PG) were assessed in terms of their impact on the innate immunity and disease resistance of Pacific white shrimp,Litopenaeus vannamei.PG derived from Bifidobacterium thermophilum was prepared in the laboratory and processed with lysozyme and protease under varying conditions to produce several different PG preparations.A standard shrimp feed was mixed with 0.05% PG preparations to produce a number of experimental diets for shrimp.The composition,concentration,and molecular weight ranges of the soluble PG were analyzed.Serum phenoloxidase and acid phosphatase activity in the shrimp were determined on Days 6-31 of the experiment.The protective activity of the PG preparations was evaluated by exposing shrimp to white spot syndrome virus(WSSV).Data on the composition of the PG preparations indicated that preparations hydrolyzed with lysozyme for 72 h had more low-molecular-weight PG than those treated for 24 h,and hydrolysis by protease enhanced efficiency of hydrolysis compared to lysozyme.SDS-PAGE showed changes in the molecular weight of the soluble PG produced by the different hydrolysis methods.Measurements of serum phenoloxidase and acid phosphatase activity levels in the shrimp indicated that the PG preparations processed with enzymes were superior to the preparation which had not undergone hydrolysis in enhancing the activity of the two serum enzymes.In addition,the preparation containing more low-molecular-weight PG enhanced the resistance of the shrimp to WSSV,whereas no increased resistance was observed for preparations containing less low-molecular-weight PG.These findings suggest that the immunity-enhancing activity of PG is related to its molecular weight and that increasing the quantity of low-molecular-weight PG can fortify the effect of immunity enhancement.     

Construction and screening of a functional metagenomic library to identify novel enzymes produced by Antarctic bacteria

A metagenomic fosmid library of approximately 52 000 clones was constructed to identify functional genes encoding cold-adapted enzymes. Metagenomic DNA was extracted from a sample of glacial meltwater,...     

Distribution of methyl mercury in <Emphasis Type="Italic">Rana chensinensis</Emphasis> and environmental media in gold-mining areas of upper reaches of Songhua River,China

The distribution characteristics of methyl mercury in Rana chensinensis and water, sediment and soil in gold-mining areas of the upper reaches of the Songhua River, China were studied by field sampling and laboratory testing. The results show that the methyl mercury contents in water, sediment and soil in gold-mining areas are much higher than those in the control site, indicating that gold-mining activities intensify the methyl mercury pollution in the study area. Methyl mercury contents are in a descending order of sediment > soil > water in the environment, and in a descending order of brain > viscera > muscle > skin in Rana chensinensis. There are significant correlation between methyl mercury contents in Rana chensinensis and those in water and sediment. In particular, the methyl mercury content in the skin of Rana chensinensis is positively correlated with those in water and sediment in spring. Therefore, skin is one of main intake pathways for methyl mercury due to its high permeability.     

Identification of Antarctic culturable bacteria able to produce diverse enzymes of potential biotechnological interest

It is estimated that more than three quarters of the Earth’s biosphere is in perennially cold environments. Despite the extreme environmental conditions of desiccation and freezing, microbes can coloni...     

Partial Sequence Analysis of Mitochondrial COI Gene of the Chinese Shrimp, Fenneropenaeus Chinensis

Eight hundred and thirty eight base pair fragment of mitochondrial COI gene of wild and cultured populations (CP1, CP4, CP5 and CP6) of Fenneropenaeus chinensis was amplified and sequenced. The A, T, G and C contents of the sequence were 235bp (28.0% ),.307bp (36.6%), 138bp (16.5%) and 158bp (18.9%), respectively. Furthermore, 556bp fragment of the sequence was used to discuss the phylogenetic relationship among 14 Penaeidae species using Alpheus armillatus as the outgroup. From the molecular phylogenetic tree constructed by neighbor-joining method, we obtained three large shrimp groups: Farfantepenaeus, Litopenaeus and Fenneropenaeus group. The results also indicated that there were a closer genetic relationships between F. aztecus and F. paulensis, L. schmitti and L. setiferus, F. indicus and F. merguiensis, and the genus Farfantepenaeus was closer to Litopenaeus.     

Genetic diversity and population structure of Eleutheronema rhadinum in the East and South China Seas revealed in mitochondrial COI sequences

Eleutheronema rhadinum is a potential commercial fisheries species and is subject to intense exploitation in China. Knowledge on the population structure of E. rhadinum in Chinese coastal waters, which is important for sustainable exploitation and proper resource management, is lacking. In the present study, the genetic diversity and population structure of E. rhadinum were evaluated using a 564-base pair fragment of the mitochondrial cytochrome c oxidase subunit I （COl） gene. A total of 76 specimens were collected from three localities around the East （Qidong and Zhoushan） and South China Seas （Zhuhai）. Among these individuals, nine polymorphic sites were detected and 1 l distinct haplotypes were defined. High levels ofhaplotype diversity （h=0.759i0.035） and low levels of nucleotide diversity （re=0.001 98i0.003 26） were observed in these populations, Hierarchical analysis of molecular variance （AMOVA） indicated that 96.72% of the genetic variation occurred within the populations, whereas 3.28% occurred among populations. No significant genealogical branches or clusters were recognized on the neighbor-joining tree. Intra-group variation among populations was significant （~0~t=0.032 85, P〈0.01）. These results suggest that E. rhadinum populations in the East and South China Seas have developed divergent genetic structures. Tests of neutral evolution and mismatch distribution suggest that E. rhadinum may have experienced a population expansion. The present study provides basic information for the conservation and sustainable exploitation of this species.     

太湖水质站网数据的主成分分析应用

本文采用主成分分析方法,对太湖湖区的水质站网数据进行了定量化研究,挖掘出对于表征水质状况的主成分变量贡献较大的水质指标,并分析了影响水质的主要原因和各个主成分的环境意义,最终求得主成分的综合得分并进行分级排序,结果表明:前四个主成分已经可以综合90%以上的因子信息,完全可以取代11个指标因子进行综合分析太湖湖区的水质状况。说明主成分分析方法克服了多维指标难以综合分析的缺陷,可能是未来水质综合分析的一种重要方法之一。     

新型壳聚糖季铵盐衍生物的合成与表征

采用甲磺酸作为溶剂兼作氨基保护试剂,对壳聚糖上的羟基选择性酯化,再游离出氨基并对其氯乙酰化,最后进行吡啶成盐,得到新型水溶性的壳聚糖季铵盐衍生物,产物的结构通过核磁共振氢谱和红外光谱进行了表征,从而为壳聚糖季铵盐衍生物的制备提供了一种新途径。     

A new identification method for five species of oysters in genus Crassostrea from China based on high-resolution melting analysis

The high phenotypic plasticity in the shell of oysters presents a challenge during taxonomic and phylogenetie studies of these economically important bivalves. However, because DNA can exhibit marked differences among morphologically similar species, DNA barcoding offers a potential means for oyster identification. We analyzed the complete sequences of the cytochrome oxidase subunit I （COI） of five common Crassostrea species in China （including Hong Kong oyster C. hongkongensis, Jinjiang oyster C. ariakensis, Portuguese oyster C. angulata, Kumamoto oyster C. sikamea, and Pacific oyster C. gigas） and screened for distinct fragments. Using these distinct fragments on a high-resolution melting analysis platform, we developed an identification method that does not rely on species-specific PCR or fragment length polymorphism and is efficient, reliable, and easy to visualize. Using a single pair of primers （Oyster- COI-1）, we were able to successfully distinguish among the five oyster species. This new method provides a simple and powerful tool for the identification of oyster species.     

A new record of Sebastes koreanus from China based on morphological characters and DNA barcoding

A new record of Sebastes koreanus(Kim and Lee,1994) was documented based on morphological characters and DNA barcoding. Fifty-six S ebastes specimens were collected from the coastal waters of northern China. Samples were identified as S. koreanus based on morphological characters. The coloration and morphometric measurements were consistent with those described from specimens collected in South Korea. In this study,specimens had the following morphological characteristics:light brown body with dark stripes and tiny dark spots,4–5 wide indistinct vertical patterns on the side,2 radial stripes behind and below the eyes,1 large dark blotch on the opercle. Additionally,the following meristic characters were recorded:dorsal fin XIV-13,pectoral fin 16,anal fin III-6–7,pelvic fin I-5,lateral line scales 29–30,and vertebrae 26. The fragment of cytochrome oxidase subunit I(C OI) gene of mitochondrial DNA was sequenced for phylogenetic analysis. The mean genetic distance within the species was 0.3%. Net genetic distances between S. koreanus and other S ebastes species ranged from 3.1% to 7.6%,which was greater than the threshold for species delimitation. The phylogenetic analysis strongly supports the validity of S. koreanus in China at the genetic level. The origion,evolution,patterns of speciation and unique features in genome divergence among primate lineages of this species still need future directions of research.     

Partial sequence analysis of mitochondrial COI gene of the chinese shrimp, <Emphasis Type="Italic">fenneropenaeus chinensis</Emphasis>

Eight hundred and thirty eight base pair fragment of mitochondrial COI gene of wild and cultured populations (CP1, CP4, CP5 and CP6) of Fenneropenaeus chinensis was amplified and sequenced. The A, T, G and C contents of the sequence were 235 bp (28.0%), 307 bp (36.6%), 138 bp (16.5%) and 158 bp (18.9%), respectively. Furthermore, 556 bp fragment of the sequence was used to discuss the phylogenetic relationship among 14 Penaeidae species using Alpheus armillatus as the outgroup. From the molecular phylogenetic tree constructed by neighbor-joining method, we obtained three large shrimp groups: Farfantepenaeus, Litopenaeus and Fenneropenaeus group. The results also indicated that there were a closer genetic relationships between F. aztecus and F. paulensis, L. schmitti and L. setiferus, F. indicus and F. merguiensis, and the genus Farfantepenaeus was closer to Litopenaeus.     

DNA barcode assessment of Ceramiales (Rhodophyta) in the intertidal zone of the northwestern Yellow Sea

A total of 142 specimens of Ceramiales(Rhodophyta) were collected each month from October 2011 to November 2012 in the intertidal zone of the northwestern Yellow Sea. These specimens covered 21 species,14 genera,and four families. Cluster analyses show that the specimens had a high diversity for the three DNA markers,namely,partial large subunit r RNA gene(LSU),universal plastid amplicon(UPA),and partial mitochondrial cytochrome c oxidase subunit I gene(COI). No intraspecific divergence was found in our collection for these markers,except for a 1–3 bp divergence in the COI of Ceramium kondoi,Symphyocladia latiuscula,and Neosiphonia japonica. Because short DNA markers were used,the phylogenetic relationships of higher taxonomic levels were hard to evaluate with poor branch support. More than half species of our collection failed to find their matched sequences owing to shortage information of DNA barcodes for macroalgae in Gen Bank or BOLD(Barcode of Life Data) Systems. Three specimens were presumed as H eterosiphonia crispella by cluster analyses on DNA barcodes assisted by morphological identification,which was the first record in the investigated area,implying that it might be a cryptic or invasive species in the coastal area of northwestern Yellow Sea. In the neighbor-joining trees of all three DNA markers,H eterosiphonia japonica converged with D asya spp. and was distant from the other Heterosiphonia spp.,implying that H. japonica had affinities to the genus Dasya. The LSU and UPA markers amplified and sequenced easier than the COI marker across the Ceramiales species,but the COI had a higher ability to discriminate between species.     

Landscape ecology: Coupling of pattern,process, and scale

Landscape ecology provides new theoretical frameworks and methodologies for understanding complex ecological phenomena at multiple scales.Studies of landscape ecology focus on understanding the dynamics of eco-logical patterns and processes,and highlight the integration of multiple disciplines.In this paper,we discussed the problems and challenges that landscape ecology is currently facing,emphasizing the limitations of current methods used to describe dynamic landscape patterns and processes.We suggested that the focus should be on the integration of ground-based observation,mobile monitoring,transect survey,and remote-sensing monitoring,as well as improved coupling of experimental and model simulations.In addition,we outlined the research frontiers in landscape ecology,including scaling,integrated pattern and process modeling,and regional synthesis.Lastly,a brief review of pat-tern-process-scale coupling studies in China was provided.We concluded by pointing out that pattern-process-scale interactions,correlations between natural,economic,and social processes,and the coupling of human and natural systems will be major research areas in landscape ecology in the future.