产甲壳素酶菌株的发酵条件、酶的分离纯化及酶学性质研究

利用甲壳素为唯一碳源从土壤中筛选得到1株产甲壳素酶活力较高的菌株HD002,初步鉴定其为Massilia属。确定菌株产甲壳素酶的最适培养基组成为(g/L):(NH4)2SO45,K2HPO40.7,KH2PO40.3,MgSO4.7H2O 1,胶体甲壳素10;最适产酶培养条件为:培养基起始pH值6.0,培养时间192 h,发酵液酶活力达1.314 U/mL。采用70%饱和度硫酸铵盐析、DEAE-Sepharose F.F阴离子交换层析、Sephadex G-75凝胶过滤层析对该甲壳素酶进行纯化,SDS-PAGE证明达到电泳纯。该甲壳素酶的分子量为60.2 kDa,最适反应温度为55℃,最适反应pH值为5.0,Cu2+和Fe3+对酶活力有明显的抑制作用,Ca2+和Na+对酶活力有明显的促进作用。     

产甲壳素酶菌株HD001的发酵条件研究及酶的分离纯化

通过对降解甲壳素菌株 HD001 的发酵条件研究,确定了其产甲壳素酶最适培养基组分为 ( w/v ):( NH4 ) 2SO4 2.0%, NaCl 0.5%, K2HPO4 0.07%, KH2P O4 0.03%, MgSO4·7H2O 0.05% 以及葡萄糖 0.1%、酵母粉 0.3% 和胶体甲壳素 1.5%;最适产酶培养条件为:培养基起始 pH 值 6.0,培养温度 30 ℃,培养时间 120 h,发酵液酶活力达 376.2 U/mL.采用硫酸铵分级盐析、DEAE-纤维素离子交换层析、Sephacryl S-100 凝胶过滤层析对该酶进行纯化后,SDS-PAGE电泳显示单一条带,其相对分子质量约为 85.7 kDa.     

硫酸软骨素酶高产菌株的筛选、鉴定和发酵培养条件的研究

通过透明圈法从土样中初筛到2株产硫酸软骨素酶能力较高的菌株,经复筛,28号菌株产酶能力更强,对该菌株的形态特征和生理生化特性考察,初步鉴定为少动鞘氨醇单胞菌(Sphingomonas paucimobilis)。该菌株的发酵产酶条件研究表明,其产酶最适培养基组分为(%,w/v):酵母粉1.0,硫酸软骨素0.5,NaCl 0.5,KH2PO40.03,MgSO4.7H2O0.5,起始pH=8.5,其余为水。其最适产酶条件为:250mL三角瓶装液量40mL,1%(v/v)接种量,25℃,100r/min培养36h。在最适产酶培养条件下,菌株发酵液中硫酸软骨素酶的活力可达1.2U/mL。本研究筛选出产硫酸软骨素酶高的菌株,并为发酵法制备硫酸软骨素酶提供一定的实验依据。     

海洋弧菌QJH-12发酵产琼胶酶条件的优化

从中国南海海域分离到了一株具有较高琼胶降解活性的弧菌Vibrio sp.QJH-12,该菌株为革兰氏阴性菌,呈弯曲杆状,极生单根鞭毛,氧化酶阳性,过氧化氢酶阳性,O/129试验阳性。在琼胶平板上生长可软化琼胶,并在菌落周围产生凹陷。通过单因子实验和正交实验确定了培养基的最佳组成:琼胶2.0‰,柠檬酸三铵1.0‰,K2HPO41.0‰,NaCl1.0%。确定该菌株的最适发酵产酶条件:装液量为250mL的三角瓶中装入125mL的液体发酵培养基,培养基起始pH6.5,摇瓶转速150r/min,30℃发酵培养26h,发酵液的酶活力达到332U/mL。结果表明,该菌株从产酶活力和发酵特点等各方面具备工业化开发的潜力。     

高产壳聚糖酶菌株的筛选和发酵产酶条件研究

为探讨专一性地降解甲壳质或壳聚糖的甲壳质酶或壳聚糖酶 ,从采集的土样中分离到 4株产壳聚糖酶能力较强的菌株 ,经摇瓶复筛 ,菌株YJ0 2产酶能力最强。对其发酵产酶条件的研究结果表明其产酶最适培养基组分为 (% ,w/v) :粉末壳聚糖 2 .0 ,葡萄糖 0 .1,NH4NO3 1.0 ,酵母提取物 0 .5 ,K2 HPO40 .0 7,KH2 PO40 .0 3 ,NaCl0 .5 ,MgSO4·7H2 O 0 .0 5 ,起始 pH 6.0。最适产酶培养条件是 :5 0 0mL三角烧瓶装瓶量为 15 0mL ,2 .0 % (v/v)接种量 ,3 0℃ ,15 0r/min培养 72h。在最适产酶培养条件下 ,72h时菌株YJ0 2发酵液中壳聚糖酶活力可达到 17.0 4U/mL。     

褐藻胶裂解酶产生菌Vibro sp.QY102的发酵条件优化

对海洋来源的Vibro sp.QY102的产褐藻胶裂解酶的发酵条件进行研究。结果表明,该菌株最适液体培养基成分为(w/v):0.5%褐藻酸钠;0.4%蛋白胨;0.3%KH2PO4;0.7%K2HPO4.3H2O;2%NaCl;0.01%MgSO4.7H2O,pH=6.0。按3%的接种量接入培养基,30℃150 r/min振荡培养120 h,产酶达到10.2 U/mL,为优化前的4.5倍。Mg2 是该菌株产酶所必需的,这在其他褐藻胶裂解酶生产菌株中未见报道。该菌株产酶发酵条件的研究,为褐藻胶裂解酶的大规模制备及应用奠定了基础。     

褐藻胶裂解酶产生菌Vibro sp．OY102的发酵条件优化

对海洋来源的Vibro sp．QY102的产褐藻胶裂解酶的发酵条件进行研究。结果表明，该菌株最适液体培养基成分为（w／v）：0．5％褐藻酸钠；0．4％蛋白胨；0．3％KH2PO4；0．7％K2HPO4·3H2O；2％NaCl；0．01％MgSO4·7H2O，pH=6．0。按3％的接种量接入培养基，30℃150r／min振荡培养120h，产酶达到10．2u／mL，为优化前的4．5倍。Mg^2＋是该菌株产酶所必需的，这在其他褐藻胶裂解酶生产菌株中未见报道。该菌株产酶发酵条件的研究，为褐藻胶裂解酶的大规模制备及应用奠定了基础。     

壳聚糖酶高产菌株的筛选和发酵条件的研究

从采集的土样中分离到1株产壳聚糖酶能力较强的菌株OU01,并对OU01进行了16S rDNA序列、形态及生理生化鉴定分析。16S rDNA序列分析表明该菌属于微杆菌属;该菌的形态特征与生理生化鉴定结果表明该菌与Microbacteriumsp.的相似性最高;因此将其初步鉴定为微杆菌属(Microbacterium)。同时对该菌的发酵条件进行了初步研究:该菌的最适培养基组成为(g/L):chitosan 10,(NH4)2SO420,MgSO4.7H2O 1.3,K2HPO4.3H2O 1.4,Glucose 1,Yeast extract 3,NaCl 5,起始pH=6.30。最适发酵温度为30℃,最佳发酵时间为96 h,在上述最优条件下,该菌株产酶达到118 U/mL。Microbacteri-umsp.OU01菌株为壳聚糖酶的研究和应用提供了新的来源。     

一株高效褐藻酸降解菌的筛选、鉴定及其发酵条件的优化

以褐藻酸钠为唯一碳源,从腐烂海带中筛选得到褐藻酸钠降解能力较强的菌株H4,经生理生化和16S rDNA序列分析鉴定该菌株属于交替单胞菌属(Alteromonas)。对菌株H4的发酵条件优化研究表明,H4的较优培养基组成为(w/v):褐藻酸钠0.6%、酵母粉0.5%、蛋白胨0.25%、NaCl 3%;较优培养条件为:培养温度28℃,初始pH7.5,接种量1.5%(v/v),培养时间48 h。Fe3+对交替单胞菌H4的产酶有明显的促进作用,这在其他褐藻酸裂解酶生产菌株中未见报道,而Cu2+对褐藻酸裂解酶的抑制作用高达45.78%。在优化后的培养条件下,粗酶液酶活达到146.45 U/mL,较优化前提高了39.4%。     

热稳定κ-卡拉胶酶产生菌Bacillus sp. Car19的发酵条件优化

对一株分泌热稳定κ-卡拉胶酶印尼热泉菌进行了种属鉴定, 并采用响应面法对该菌发酵产酶条件进行了优化。鉴定结果表明, 该菌株属于芽孢杆菌属(Bacillus), 命名为Bacillus sp. Car19(GeneBank: KT865196)。发酵条件优化结果显示, 9个环境因子影响Bacillus sp. Car19产酶量。其中影响Bacillus sp. Car19产酶量的三个主要因素分别为培养温度、培养基中Cu²⁺浓度和培养基中NaCl浓度。综合次要因素对Bacillus sp. Car19产酶影响, Bacillus sp. Car19最佳产酶发酵条件为: 培养温度52.31℃、Cu²⁺浓度6.93 mmol/L、NaCl浓度37.03 g/L, 培养基pH为6, 接种量1%, 培养时间36h, 半乳糖浓度0.3 g/L,硝酸铵浓度7g/L, 卡拉胶浓度0.5g/L。优化后发酵上清液酶活力达到15.21 U/mL, 与优化前相比提高了1.5 倍。     

Model of the post-Cenozoic evolution of the cryolithozone of the shelf of the western part of the Laptev Sea

The analysis of the structure of the cryolithozone, facies, and thicknesses of the Quaternary sediments and the results of the physicochemical mathematical modeling of the modern shelf of the western part of the Laptev Sea support the influence of the Late Pleistocene glaciations on the heat conditions and the distribution of the permafrost in the area. A ～200-m thick glacier formed under aerial conditions from atmospheric precipitation represented the metamorphosed snow cover. According to the modeling, the long-living (from 60?50 to 10?4 ky) glacier reduced the thickness of the permafrost rocks in the reviewed shelf area for 280–360 m. The Holocene marine transgression additionally decreased the thickness from 50–140 m on the inner shelf to 220–350 m on the outer shelf. The modern submarine cryolithozone 450-0 m thick is wide-spread in the studied region from the coast to the shelf boundary (isobaths of 130–140 m), where it pinches out at a distance of ～380 km from the coast at a depth of ～250 m above the sea level.     

Investigation of the mechanism of the long-term evolution of the carbon cycle in the ecosystem of the Sevastopol bay

We analyze the components of the carbon system of the Sevastopol bay waters and the balance of main sediment-forming substances using the data of field investigations in 1998–2008. The interannual variations of total inorganic carbon and the equilibrium partial pressure of carbon dioxide in the bay water are noted. An increase in the flux of carbon dioxide into the bay and in the content of organic carbon in bottom sediments is revealed, and an explanation of this phenomenon is given. The priority accumulation of organic carbon in the sediments of the bay is established. We assess the interannual variation in the relative abundances of organic and inorganic carbon as an index of the carbon cycle stability.     

Contemporary state of the investigation of the influence of the discharge of rivers on the hydrologic structure of the Black Sea

We generalize and analyze the state of investigations of the influence of river waters on the hydrologic structure of the Black Sea. Specific features of the water, regime and hydrography of rivers flowing into the sea, data on the discharge of rivers, and intrayear and interyear variability of the discharge are presented. We discuss and analyze works studying the transformation of river waters, routes of desalinized sea waters, and their influence on the thermohaline structure and dynamic regime of the sea. Translated by Peter V. Malyshev and Dmitry V. Malyshev     

双壳贝类血淋巴中儿茶酚胺的检测方法初步研究

研究的目的是探索一种检测双壳贝类血淋巴中儿茶酚胺含量的方法。以栉孔扇贝(Chlamysfarreri)和长牡蛎(Crassostreagigas)为实验对象,研究了取样、抗氧化剂、氧化铝用量以及Tris缓冲液用量等对实验结果的影响。结果表明,高效液相色谱电化学检测器法(HPLC-ECD)可以灵敏高效地对双壳贝类血淋巴中儿茶酚胺进行定性、定量检测。实验表明,取样时间最长不超过1.5min;还原性谷胱甘肽作抗氧化剂效果较好;前处理各试剂与血样量的最适配比为血样量1.5mL,Tris缓冲液(1.5mol/L,pH8.6)1mL和氧化铝25mg;水洗后离心并尽可能吸干氧化铝中的水分。去甲肾上腺素、肾上腺素和多巴胺的回收率分别为53%~69%、47%~73%和48%~56%。     

黄河入海径流对河口海域盐度影响研究

为研究黄河入海径流变化条件下河口附近海域盐度扩散特征,以更好地保护河口海域生物资源多样性,本文以黄河下游利津水文站的长序列实测径流数据资料为基础,利用近海水动力模型FVCOM,分析径流变化对黄河口海域盐度的影响规律。结果表明：黄河口与莱州湾之间存在顺时针的环流系统,在余流作用以及涨落潮方向的影响下,黄河冲淡水长期向莱州湾扩散;丰水期黄河冲淡水几乎影响了整个莱州湾,27盐度锋可以到达莱州湾中部,27等盐线的表层包络面积为2 665.61 km²,占莱州湾的1/4左右,枯水期低盐度水只有向南扩散的趋势,27以下的低盐度水集中分布在黄河口门附近,27等盐线的表层包络面积只有199.65 km²;5月份,随着入海径流量增加,27等盐线扩散的范围、距离、方向都会发生明显变化。在对近海生物资源有迫切保护需求的情景下,适当减少其他用水户供水量以增加入海生态径流量,可以有效改善黄河口海域附近的盐度情况,为生物资源的生长繁殖创造良好条件。     

Modelling of the ecosystem of the Black-Sea shelf zone near the estuary of the Danube

We analyze the results of numerical calculations performed according to the three-dimensional interdisciplinary model of an ecological system of the Black-Sea shelf zone near the estuary of the Danube. The complete system of equations of hydrothermodynamics is solved together with transport equations of the advection-diffusion-reaction type used to describe the transformation of a substance (nitrogen) between the components of the characteristic vectors of the ecosystem: plankton, detritus, and biogenic elements (nitrates). We describe the distinctive features of the circumcontinental distribution of components obtained as a result of numerical experiments and present arguments for the conclusion that the ecosystem of the Danube estuary water area plays the role of a buffer zone between the press of the Danubian biogenic pollutions and the neighbouring areas of the shelf zone and open sea. Translated by Peter V. Malyshev and Dmitry V. Malyshev     

The influence of the bottom contour on the deformed state of the ice cover due to the motion of the submarine

The authors have previously determined that the effectiveness and failure pattern of the ice cover caused by flexural-gravity waves generated by a submerged body motion near the bottom ice can greatly depend on the depth of the water area. In its turn, the presence of a ledge on the ice surface may affect a wave propagation pattern. This paper presents an experimental study of the bottom contour influence on the deflection and length of flexural-gravity waves. The authors describe a numerical model for the analysis of the deformed state of ice caused by hydrodynamic loads due to a submarine motion, taking into account the bottom contour. The experiments are carried out in the ice tank. The results of calculations and experiments are compared.     

Dynamics of the ice cover and peculiarities of the ice transportation of the sediments at the tidal flats of the Kandalaksha Gulf of the White Sea

Ten years of semistationary studies of the interaction of the ice cover and the coastal relief of the Kandalakhsa Gulf in the White Sea allowed us to reveal the peculiarities of these processes in the warm and cold years and the mechanisms of the origination of ice forms, including ice tents, which govern the character of the tidal flats. Three dynamic fast ice zones are distinguished by their geomorphological location, morphology, and ice dynamics. The dependence of the mechanisms and volumes of the enrichment of the fast ice in the sediments on the coastal zone??s relief was established. At the coast of the Kindo Peninsula (Velikaya Salma Strait), the ice mostly contains fine-sandy sediments. Coarser sediments, including boulders, are occasional and frozen into the ice at the contact of the fast ice and the bottom. At the expense of the ice??s spreading by 4?C6 times in the warm years and 2?C4 times in the cold years, several thousand tons of sediments mostly from the tidal flat??s surface (100?C700 m³ from 1 km of coast in the wide tidal flats and <50 m³ on the steep coasts) are delivered to the water area.     

Relationship between the path of the Kuroshio in the south of Japan and the path of the Kuroshio Extension in the east

A relationship between paths of the Kuroshio and Kuroshio Extension (KE) is investigated, using the satellite-derived altimetry dataset of 1993–2008. When the Kuroshio takes the nearshore nonlarge meander path or typical large meander path and resultantly goes through the deeper channel (about 2500 m) of the Izu-Ogasawara Ridge, the KE path adopts a relatively stable state with the two quasi-stationary meanders. On the other hand, when the Kuroshio takes the offshore nonlarge meander path and then passes over the shallower part of the Ridge (about 1000 m), the KE path tends to be convoluted, i.e., an unstable state.     

A model of the dynamics of the ecosystem of the Sea of Azov

We use many-year archival observations of hydrologic and hydrobiologic state of the Sea of Azov for the identification of the structure of a model of its ecosystem. The ecosystem model, supplemented with expert estimates of bioresource consumption (fish reserve), contamination level, and possible ecologic fines for violation of the sea natural state, is formalized by the method of system dynamics. Hereat, the major part of influence functions is found in terms of observational data with application of self-organization algorithms. We also present the results of simulated experiments with the model of the ecosystem, which enable us to analyse scenarios of its behaviour under the influence of various external factors (wind, river discharge, water exchange with the Black Sea, etc.). Translated by Peter V. Malyshev and Dmitry V. Malyshev