Construction of recombinant Pichia pastoris for β-1，3-1，4-glucanase expression and optimization of fermentation process for glucanase production

β-1,3-1,4-葡聚糖酶(EC3.2.1.73)作为一种重要的工业用酶，被广泛应用于酿酒和饲料工业中。目前国内对葡聚糖酶的研究还处于初级阶段，天然菌株产酶量及酶性能满足不了实际应用。随着基因工程方法的应用，为实现目的蛋白的工程菌表达提供的了可能。现国内外对β-1,3-1,4-葡聚糖酶的研究集中在构建工程菌株来提高酶的产量和性能方面。本研究的主要目的即通过基因工程及分子生物学方法来提高β-1,3-1,4-葡聚糖酶表达量，研究其酶学特性，为生产中的问题提供有效的解决途径。主要研究内容及相应结果如下： 采用PCR法从载体pLF3中扩增出杂合β-1,3-1,4-葡聚糖酶基因，并在基因两端引入酶切...β-1,3-1,4-glucanase(EC 3.2.1.73), as an important industrial enzyme，has been widely used in the brewing and animal feed industry. Nowadays the domestic research on β-1,3-1,4-glucanase is in the beginning, and the expression level and properties of natural enzyme are not sufficient for industrial purpose. The development of molecular biology and genetic engineering provides a feasible way to resolv...学位：工学硕士院系专业：化学化工学院化学工程与生物工程系_生物化工学号：2062006115203

Observation of Annual Growth of Two Sponge Explants

为了解决药源海绵的供给问题,促进活性产物的挖掘和筛选,针对福建近海2种海绵:叶片山海绵(Mycale phyllophila)和苔海绵(Tedania sp.)的周年生长率以及生长模式开展了系统研究,旨在为实现药源海绵的规模化移植提供必要的实践依据.采用原位尼龙绳挂养的方式,对随机挑选的各30株海绵,每月进行生长数据的测量,并对海区水温进行周年高密度记录.其中苔海绵的观察时间为2012年7月至2013年8月,叶片山海绵的观察时间为2013年6月至2014年4月.结果表明:叶片山海绵的生长率与水温变化显著相关(p0.05),在2013年8月出现最高周长生长率,增长率为93.88%;长度年增长率为(546.45±201.98)%,最大的个体长度增长率出现在8月,达到了956.11%.2种海绵的生长模式相似,均是优先覆盖尼龙绳等附着基的表面,之后才开始向着增加厚度的方向发展.根据实验结果,在海区逐步出现海绵的季节,海绵的移植应该选择在水温开始升高的4—5月,叶片山海绵采用固定在绳索上的移植方式,苔海绵则采用网兜移植的方式,有望获得较理想的移植效果.In order to solve the supply shortage of pharmaceutical sponges as well as to promote mining of active products and selection screening,our research focuses on studying the annual growth rate and growth pattern of two sponges,Mycale phyllophila and Tedania sp.,which are common species in coastal waters of Fujian.The aim of this study is to provide practical basis for achieving the scale transplantation of pharmaceutical sponge.We applied an approach of aquaculture in situ using nylon ropes and randomly picked up 30 individuals of sponges for cultivation.We recorded the growth data of every single individual,and the temperature and light automatic recorder was used to record the sea water temperature during the experiment.The experiment on Tedania sp. lasted for more than 12 months from July 2012 to August 2013,while the experiment on M. phyllophila lasted for about one year from June 2013 to April 2014.The result showed that the growth rate of M.phyllophila correlated significantly with sea water temperature( p 0. 05). In August 2013,they grew fastest with the peak perimeter growth rate of93. 88%.The length growth rate was( 546. 45±201. 98) %,and the largest individual had a length growth rate of 956. 11%. The growth strategies of these two species are similar,and they prefer to occupying more growing space by covering the substrate before expanding their thickness.According to the results,we suggest that transplanting experiment on sponges should be done around April to May when the water temperature is warm enough for sponges to survive.For M. phyllophila,ropes are better substrate for transplanted individuals to grow while string bags are expected to be effective for Tedania sp. to survive.厦门南方海洋研究中心项目(13GYY002NF07

智能制造刀具管理系统及刀具剩余寿命监测功能开发

伴随智能制造技术的深入推进,刀具管理系统的功能根据用户业务需求在不断拓展。针对工艺设计后的刀具选购、使用直至报废等环节,基于\"刀具流\"开发了一款智能制造刀具管理系统,更精确地实现了刀具使用过程的数字化、网络化、智能化和可视化全生命周期管理。鉴于目前刀具状态监测技术的复杂和可靠性低等问题,在工程应用上提出了一种基于被加工工件表面粗糙度间接预测刀具剩余寿命的方法。整个系统在现场得到了良好应用。2016年工信部智能制造综合标准化与新模式应用项目(工信部联装[2016]213号);;国家自然科学基金项目(51605403);;福建省产业技术联合创新专项(闽发改投资[2016]482号

Clone and expression of β-1,3-1,4-glucanase in Pichia pastoris

β-1,3-1,4-d-葡聚糖酶是一类专一性降解β-1,3-1,4-葡糖苷键中的β-1,4-糖苷键,产生小分子还原糖的水解酶,广泛应用于啤酒工业和饲料工业中。本研究根据毕赤酵母密码子偏好性优化β-1,3-1,4-葡聚糖酶基因序列,采用PCr法将其插入毕赤酵母表达载体PPICzαA,经SACI线性化后电击整合入毕赤酵母X-33基因组,构建重组酵母;经菌落PCr验证和摇瓶筛选,获得一株X-33/PPICzαA-bgl,甲醇诱导96H后,酶活力达308.5u/Ml,经SdS-PAgE电泳,实际蛋白分子量约为33ku。β-1,3-1,4-d-葡聚糖酶最适反应PH为5.0,最适反应温度为50℃。β-1,3-1,4-glucanase,which can hydrolyzeβ-1,3-1,4-glucan by cutting off theβ-1,4-glycosidic linkages and release shorter reducing sugar,has been applied in the brewing and animal feed additive industry.It can effectively improve digestibility of barley-based diets and reduce enteritis.In this report,the gene ofβ-1,3-1,4-glucanase,which was optimized according to Pichia pastoris condon usage,was amplified by polymerase chain reaction and ligated into the expression vector pPICZαA.The recombinant vector was lineared by Sac I and transformed into Pichia pastoris X-33 by electroporation.Then the optimal positive transformant,named X-33/pPICZαA-bgl,was screened by colony PCR and shake flask fermentation.The enzyme activity of recombinantβ-1,3-1,4-glucanase couldreach 308.5U/mL after 96h methanol induction.Results of SDS-PAGE showed that about 33ku protein of theβ-1,3-1,4-glucanase was expressed.Acidity and temperature optimal for this recombinant enzyme was pH5.0 and 50℃,respectively.福建省农业科学院青年基金(2012DQC-11); 公益性行业(农业)科研专项(201303094-05

海西对台光电产业产学研合作现状及趋势

海峡西岸对台光电产业产学研合作是两岸科技界高度重视的一项重点工程,在两岸政策的引导下,在市场需求及产业界、学术界的共同推动下,合作领域越发广泛,合作交流的形式和渠道不断拓宽,带动两岸科技进步及经济发展。本文着重分析了两岸光电产业的发展现状和海西对台光电产业产学研合作现状、趋势,阐明了合作机制及相应的目标与重点、方式等。在此基础上提出了海峡西岸经济区对台光电产业合作的策略及对两岸光电产业良性发展的建议。福建省科技计划项目（2016R0091）;福建省高等教育教学改革研究项目（JAS151398）

Determination of Pseudostellarin B in Pseudostellaria heterophylla by HPCE and Study on Its Fingerprints Analysis

建立了测定太子参中环肽Pseudostellarin B含量的高效毛细管电泳(HPCE)分析方法,对10种不同产地太子参中环肽Pseudostellarin B的含量进行了测定。HPCE工作条件:采用未涂层熔融石英毛细管(内径75μm,有效长度50cm),分离电压为15kV,柱温20℃,二极管阵列检测器(DAD)检测波长为203nm,缓冲液为20mmol/L硼砂(pH=9.3)溶液。在选定的工作条件下,环肽Pseudostellarin B浓度与其响应信号值之间具有较好的线性相关性(Y=0.6357X+2.546,R=0.9985),加标回收率在93.8%~105.6%之间。在此基础上进行了太子参HPCE指纹图谱研究,采用中药指纹图谱相似度计算软件对不同太子参样品的HPCE指纹图谱进行相似度计算,以系统生成的对照指纹图谱为对照模板,10份样品中有8份的相似度在0.90以上,说明该方法可用于太子参质量控制。A modern HPCE analysis method was developed for determinating cyclic peptide Pseudostellarin B in Pseudostellaria heterophylla(Miq.) Pax.Separation and determination were carried out by HPCE under the following conditions: bared fused silica capillary (50 cm×75 μm i.d.),20 mmol/L borate (pH=9.3)as buffer,the run voltage is +15 kV,detection length of UV at 203 nm,and column temperature of 20 ℃.The results indicated that the developed method was simple,accurate and reliable for the determination of Pseudostellarin B with a good linearity(Y=0.6357X+2.546,R=0.9985),and the recovery ranges of 93.8%～105.6%.Based on this method,the HPCE fingerprints of Pseudostellaria heterophylla (Miq.) Pax were established.The fingerprints were compared by the software of the similarity evaluation system for chromatographic fingerprint.The fingerprint congruence coefficients of 8 electropherograms in 10 were above 0.90.The method of HPCE fingerprint is reliable and accurate.国家自然科学基金项目(20235020);; 国家科技部攻关项目(2003BA759C

Analysis on death causes of liver cancer of inhabitants in Xiamen City from 2002 to 2011

目的了解2002-2011年厦门市居民肝癌死亡趋势、分布特征及寿命损失情况。方法对2002-2011年厦门市肝癌死亡登记资料和人口数据进行统计分析,通过计算死亡率、标化死亡率、年均增长死亡率及寿命损失年等指标,评价居民的肝癌死亡及寿命损失情况。结果 2002-2011年厦门市肝癌年均死亡率为32.70/10万,标化死亡率为34.48/10万,居恶性肿瘤死亡顺位第1位,总体趋势有下降,但差异无统计学意义(2趋势=2.13,P=0.144)。男性、女性、城区、郊区、农村的肝癌年均死亡率均呈下降趋势,但只有郊区下降有统计学意义(2趋势=7.46,P=0.006)。肝癌死亡率比较:农村>城区>郊区,男性死亡率高于女性,差异有统计学意义(2=1 758.80,P<0.001)。肝癌主要危害中老年人群,死亡年龄的中位数为57岁,89.37%的死亡病例发生在40岁以上人群。肝癌潜在寿命损失年为93 378.0人年,平均减寿年数为20.17人年,减寿率为5.74‰,男性和农村居民寿命损失较严重。结论 2002-2011年厦门市肝癌死亡率整体呈下降趋势,但对中老年人群的危害仍很严重,应继续加强防控力度。Objective To gain the knowledge of trends in mortality,distribution characteristics and potential years of life lost caused by liver cancer among Xiamen population from 2002 to 2011.Methods Mortality rate,standardized mortality rate,average annual growth rate of mortality and years of life lost have been employed to evaluate the mortality of liver cancer and years of life lost caused by the disease among population in Xiamen.Results The average annual mortality rate of liver cancer among Xiamen population was 32.70 /105.After the standardization,the rate was ascended to 34.48 /105,ranking in the top of the death causes.Compared to historical data,the mortality rate slightly decreased in total.However,no statistical significance was observed.The average annual mortality rate of male,female,urban,suburban and rural areas were decreased,but only a statistically significant decrease in the suburban was noted.As for the locations,the mortality rate in rural area was the highest,and that of urban area maintained in the second position,and the rate of suburb area was the lowest in Xiamen.The reduction on the average mortality rate among female population was greater than that of its counterparts.We also observed that the reduction was greater in rural areas than in the rest of city.However,the differences were not statistically significant.Liver cancer mainly affected the middle-aged and old population,and the median of death age was 57 years old.Furthermore,89.37% of deaths were reported in the population older than 40 of years.The potential years of life lost caused by liver cancer among Xiamen population was 93 378.0 PY and AYLL was 20.17 PY,and PYLL rate reached 5.74‰.The years of life lost caused by liver cancer among male and rural population are serious.Conclusions The mortality rate of live cancer among Xiamen population between 2002 and 2011 has been declined in general aspect.However,the liver cancer still renders great threat to the health among middle-aged and old population.Therefore,the prevention and control strategy should be further promoted.福建省卫生厅青年科研课题(2010-2-113

Testing System of Phosphor Excited by Blue LED Based on Integral Sphere

为尽可能反映荧光粉在实际工作条件下的发光性质,准确测量荧光粉的发光性能,提出了一种基于积分球的荧光粉发光性能测试系统。该系统采用蓝光lEd作为激发光源,配置有TEC控温系统对光源有效实施控温,可提供持续稳定的激发荧光粉发光。采用出光筒控制光源方向和积分球收集光线,能有效防止能量损失,提高测试的精确性。通过实验得到了不同强度蓝光激发下的荧光粉光谱功率分布以及发光效能、量子效率、光转换效率等参数的变化规律。随着驱动电流的增大,由于蓝光芯片内量子限制斯塔克效应,从而导致蓝光峰值波长出现小幅度的蓝移。三种效率在小电流下基本呈线性下降趋势,且在大电流下趋于平缓。实验结果表明,该系统及方法可以有效地评价实际lEd芯片工作状态下的荧光粉发光性能。A testing system for phosphor luminous characteristics based on integral sphere is proposed so as to show luminous characteristics of phosphor at actual operation circumstances and measure its luminous characteristics accurately.Blue LED is used as an excited source in the system.TEC temperature control system is used to control the temperature of the source effectively.And continuous and stable excited light is provided to excite phosphor illumining.Energy loss is prevented effectively and testing accuracy is enhanced for the direction of source controlled by a light tube and light ray collected by integral sphere.Based on experiments,spectrum power distributions of phosphor excited by blue light with difference light strength and variation rules of parameters such as luminous efficiency,quantum efficiency and light conversion efficiency are got.With the increasing of driving current,a small blue shift appears in blue light peak wavelength for quantum confined Stark effect in blue light chip.Three kinds of efficiency almost show linear decline trend at low current and then approach to constant at high current.Experimental results show that luminous characteristics of phosphor on real LED chip at operation state can be evaluated effectively by the system and method.国家自然科学基金项目(11104230); 福建省重点科技项目(2011H6025;2012H0039

White LED Phosphor Performance Tested by Spectro320e Spectrometer

我国lEd产业发展非常迅速,稀土钇铝石榴石(yAg)系列荧光粉广泛应用于白光lEd。德国InSTruMEnT SySTEMS公司的SPECTrO320E光谱仪适合光谱的精确测量,文章结合SPECTrO320E和远方公司的PE-5荧光粉激发装置,实现蓝光激发下黄色yAg荧光粉的色品坐标、相对亮度、量子效率等参数精确的测试,研究了仪器在不同的测试条件如暗电流、光电倍增管、密度滤光片等设置的不同,对测试结果的影响,并给出利用本装置测试荧光粉时应该注意的事项。LED industry in china develops rapidly.Yttrium aluminitum garnet(YAG) phosphors are widely applied in white LEDs.The Instrument Systems company's Spectro320e spectrometer is suitable for accurate measurement of the spectrum.It is combined in this paper with the Everfine company's PE-5 phosphor excitation device to achieve the accurate testing of yellow YAG phosphors excited by blue light,such as chromaticity coordinates,relative brightness and quantum efficiency.We also studied the influence of different testing conditions on the results,such as dark current,PMT and density filter.The cautions are provided herewhen using the Spectro320e spectrometer

Thermal characteristic analysis of high-power LEDs by structure functions

运用电学法测量功率型lEd冷却瞬态温度曲线,通过数学方法将其转化为积分和微分结构函数来分析器件各区域的热阻和热容,结果发现,各层材料的测量值与理论值基本一致。1μS的瞬态数据采集精度和高的重复性保证了实验结果的准确性和可靠性,运用这种方法比较了3种不同金属芯印刷电路板(MCPCb)对功率型lEd的散热效果,贝格斯Al基板散热性能最好,AnTAl基板次之,普通Al基板最差。研究表明,利用结构函数分析功率型lEd的热特性是一种强有力的方法。The cooling transient temperature curves of high-power LEDs are measured by electritical method.The cumulative and differential structure functions are extracted from these curves to analyse thermal resistances and thermal capacitances of all regimes of high-power LEDs with numeric computational method.It is found out that calculated and measured values of various materials are essentially conformable.The sampling resolution of 1 μs of transient data and high repetition assure the veracity and reliability of experimental result.Subsequently,thermal conduction capabilities of three different metal core printed circuit boards(MCPCBs) with high-power LEDs are compared by this method,and it is discovered that bergquist′s MCPCB has the best thermal conduction capability,ANT′s MCPCB takes second place,and the common MCPCB is the worst.So the structure functions are powerful tools for thermal characteristic analysis of high-power LEDs.国家“863”计划资助项目(2006AA03A175);福建省科技项目(2006H0092;2008J0030);厦门市重大专项资助项目(3502Z20061004