Chelerythrine_Chloride_COA_26018_MedChemExpress
广藿香醇结构式
广藿香醇结构式全文共四篇示例，供读者参考第一篇示例：广藿香（Magnolia officinalis），又名辣藿香、辣木香、白藿香等，是一种常见的植物，属于木兰科藿香属。
广藿香主要生长在中国、日本、朝鲜和俄罗斯等地区，是一种常见的药用植物。
广藿香的主要有效成分是广藿香醇，是一种具有多种药理作用的重要化合物。
广藿香醇的结构式为C10H18O，化学名为4-allyl-1-methoxy-2-(1-propenyl)cyclohexene，是一种萜烃类化合物。
广藿香醇是广藿香中的一种活性成分，具有多种生物活性和药理作用，被广泛应用于中药制剂、保健品和化妆品中。
广藿香醇具有镇静安神、抑制细菌、抗炎以及减轻疼痛等功效。
研究表明，广藿香醇具有抗抑郁、镇静安眠、抗氧化等作用，可有效缓解精神压力，改善睡眠质量，提高身体抵抗力。
广藿香醇还具有抗菌、抗炎和抗氧化作用，可以有效预防和治疗多种疾病。
广藿香醇的应用领域非常广泛，既可以直接作为中药成分使用，也可以应用于化妆品、保健品等领域。
广藿香醇在中药领域被广泛应用于治疗失眠、焦虑、忧郁、神经衰弱等症状，其镇静安神、抗抑郁的作用备受认可。
在化妆品领域，广藿香醇具有很好的抗氧化和抗菌功效，可以用于护肤品、美容品中，帮助净化肌肤、抗衰老和延缓皮肤衰老。
广藿香醇作为广藿香中的重要有效成分，具有多种生物活性和药理作用，被广泛应用于中药制剂、保健品和化妆品中。
其镇静安神、抗氧化、抗菌等功效备受认可，有望成为未来中药和化妆品市场中的热门产品。
随着人们对健康、自然的追求，广藿香醇的市场前景将更加广阔，对于广藿香的开发利用和推广具有积极的意义。
第二篇示例：广藿香醇，又称广香藿醇，是一种具有特殊香气的有机化合物，常被用于食品、医药和香料行业。
它的独特香气使其成为一种重要的香料和药用成分，具有镇静、抗菌、抗炎和抗氧化等多种功效。
广藿香醇的化学结构式为C10H18O，是一种萜类天然产物。
广藿香醇的结构式如下：由结构式可知，广藿香醇分子由一个环烷基和一个烯醇基组成。
3-氯-2-肼基吡啶 分子量
3-氯-2-肼基吡啶分子量
3-氯-2-肼基吡啶（3-chloro-2-hydrazinylpyridine）是一种有机化合物，化学式为C5H5ClN4，分子量为158.57 g/mol。
它是一种重要的中间体化合物，在医药和农药等领域具有广泛的应用。
3-氯-2-肼基吡啶是一种白色到浅黄色的固体，具有较高的稳定性和溶解性。
它可以通过吡啶-3-羧酸和盐酸肼反应得到，也可以通过其他合成路线进行制备。
在生产过程中，需要严格控制反应条件，以保证产物的纯度和产率。
在医药领域，3-氯-2-肼基吡啶被广泛用于合成药物。
它可以作为一种重要的合成中间体，参与到许多药物的合成过程中。
比如，它可以用于合成抗癌药物、抗菌药物等。
同时，3-氯-2-肼基吡啶本身也具有一定的生物活性，可以作为一种药物原料直接用于制备药物。
在农药领域，3-氯-2-肼基吡啶也是一种重要的中间体。
它可以用于合成各种类型的农药，如杀菌剂、杀虫剂等。
这些农药在农业生产中起到了重要的作用，可以有效地控制农作物病虫害，提高农作物产量。
除了医药和农药领域，3-氯-2-肼基吡啶还可以用于其他领域。
比如，它可以用于有机合成反应中的催化剂，可以参与到材料合成中，还可以用于有机电子材料的制备等。
总的来说，3-氯-2-肼基吡啶作为一种重要的中间体化合物，具有广泛的应用前景。
随着化学合成技术的不断发展和改进，相信它将会在更多的领域展现其价值，为人们的生活和生产带来更多的便利。
【藥品名】氯丙嗪【英文名】chlorpromazine【別名】氯普馬嗪；冬眠靈
【药品名】氯丙嗪【英文名】Chlorpromazine【别名】氯普马嗪；冬眠灵；可乐静；可平静；氯硫二苯胺；Chlorpromazium；Wintermin；Aminazine【剂型】1.片剂：12.5mg，25mg，50mg；2.注射剂(粉)：10mg，25mg，50mg；3.复方制剂：复方氯丙嗪片：每片含盐酸氯丙嗪及盐酸异丙嗪各12.5mg；复方注射剂：每2ml含盐酸氯丙嗪及盐酸异丙嗪各25mg；4.冬眠合剂：盐酸氯丙嗪、盐酸异丙嗪各50mg，哌替啶100mg，加入5%葡萄糖注射剂中，静脉输注，用于冬眠疗法。
【药理作用】本药属二甲胺族吩噻嗪类药物，为抗精神病药的代表药。
主要阻断脑内多巴胺受体，这是本药抗精神病作用的机制，也是长期应用产生严重不良反应的基础。
本药还能阻断α肾上腺素受体和M胆碱受体，因而其药理作用广泛。
具体作用如下：1.抗精神病作用：目前认为，本药通过阻断与情绪和思维有关的边缘系统的多巴胺受体而起抗精神病作用。
而阻断网状结构上行激活系统的α肾上腺素受体，则与镇静安定的作用有关。
正常人服用治疗量后，出现安静、活动减少、感情淡漠、注意力降低、对周围事物不感兴趣等反应。
安静时可诱导入睡，但易被唤醒。
精神患者服用后，在不过分抑制的情况下，可迅速控制精神分裂症患者的躁狂症状，减少或消除幻觉、妄想等症状，使思维活动及行为趋于正常。
2.镇吐作用：小剂量可抑制延脑催吐化学敏感区的多巴胺受体，大剂量时又可直接抑制呕吐中枢，产生强大的镇吐作用。
但对刺激前庭所致的呕吐无效。
3.降温作用：本药对下丘脑体温调节中枢有很强的抑制作用，不但降低发热患者的体温，还能降低正常体温，这与解热镇痛药不同，后者只降低发热体温而不降低正常体温。
本药的降温作用随外界环境温度而变化，环境温度愈低其降温作用愈明显，与物理降温同时运用具有协同作用，在炎热的天气，本药反可使体温升高，这是干扰了机体正常散热的结果。
4.阻断外周肾上腺素受体，直接扩张血管，引起血压下降。
大肠杆菌高产L丙氨酸
APPLIED GENETICS AND MOLECULAR BIOTECHNOLOGYProduction of L-alanine by metabolically engineered Escherichia coliXueli Zhang&Kaemwich Jantama&J.C.Moore&K.T.Shanmugam&L.O.IngramReceived:23May2007/Revised:13August2007/Accepted:16August2007/Published online:15September2007 #Springer-Verlag2007Abstract Escherichia coli W was genetically engineered to produce L-alanine as the primary fermentation product from sugars by replacing the native D-lactate dehydroge-nase of E.coli SZ194with alanine dehydrogenase from Geobacillus stearothermophilus.As a result,the heterolo-gous alanine dehydrogenase gene was integrated under the regulation of the native D-lactate dehydrogenase(ldhA) promoter.This homologous promoter is growth-regulated and provides high levels of expression during anaerobic fermentation.Strain XZ111accumulated alanine as the primary product during glucose fermentation.The methyl-glyoxal synthase gene(mgsA)was deleted to eliminate low levels of lactate and improve growth,and the catabolic alanine racemase gene(dadX)was deleted to minimize conversion of L-alanine to D-alanine.In these strains,re-duced nicotinamide adenine dinucleotide oxidation during alanine biosynthesis is obligately linked to adenosine triphosphate production and cell growth.This linkage provided a basis for metabolic evolution where selection for improvements in growth coselected for increased glycolytic flux and alanine production.The resulting strain, XZ132,produced1,279mmol alanine from120g l−1 glucose within48h during batch fermentation in the mineral salts medium.The alanine yield was95%on a weight basis(g g−1glucose)with a chiral purity greater than99.5%L-alanine.Keywords Alanine.Fermentation.E.coli.Evolution. GlycolysisIntroductionWorldwide production of L-alanine has been estimated at 500tons per year(Ikeda2003).In pharmaceutical and veterinary applications,L-alanine is used with other L-amino acids as a pre-and postoperative nutrition therapy(Hols et al.1999).Alanine is also used as a food additive because of its sweet taste(Lee et al.2004).The use of L-alanine is limited in part by the current high cost.L-Alanine is pro-duced commercially by the enzymatic decarboxylation of L-aspartic acid using immobilized cells or cell suspensions of Pseudomonas dacunhae as a biocatalyst with a yield greater than90%(Shibatani et al.1979).The substrate for this enzymatic production process,L-aspartate,is usually pro-duced from fumarate by enzymatic catalysis with aspartate ammonia-lyase.Fumaric acid is produced primarily from petroleum,a nonrenewable feedstock.An efficient fermen-tative process with a renewable feedstock such as glucose offers the potential to reduce L-alanine cost and facilitate a broad expansion of the alanine market into other products.Alanine is a central intermediate(Fig.1)and an essential component of cellular proteins.Most microorganisms produce alanine only for biosynthesis using a glutamate–pyruvate transaminase(Hashimoto and Katsumata1998). Some organisms such as Arthrobacter oxydans(Hashimoto and Katsumata1993;Hashimoto and Katsumata1998; Hashimoto and Katsumata1999),Bacillus sphaericus (Ohashima and Soda1979),and Clostridium sp.P2Appl Microbiol Biotechnol(2007)77:355–366DOI10.1007/s00253-007-1170-yElectronic supplementary material The online version of this article (doi:10.1007/s00253-007-1170-y)contains supplementary material, which is available to authorized users.X.Zhang:J.C.Moore:K.T.Shanmugam:L.O.Ingram(*) Department of Microbiology and Cell Science,University of Florida,Box110700,Gainesville,FL32611,USAe-mail:ingram@K.JantamaDepartment of Chemical Engineering,University of Florida, Gainesville,FL32611,USA(Orlygsson et al.1995)produce alanine from pyruvate and ammonia using an reduced nicotinamide adenine dinucleo-tide (NADH)-linked alanine dehydrogenase (ALD).How-ever,fermentations are slow,and yields from the best natural producers are typically 60%or less because of coproduct formation (Hashimoto and Katsumata 1998;Table 1).Plasmid-borne genes encoding NADH-linked ALD have been tested as an approach to develop improved biocatalysts with varying degrees of success (Table 1).Engineered strains of Zymomonas mobilis CP4expressing the B.sphaericus alaD gene produced low levels of racemic alanine during the anaerobic fermentation of 5%glucose (Uhlenbusch et al.1991).A native chromosomal lactate dehydrogenase gene (ldhA )-deleted strain of Lactococcus lactis containing a mutation in alanine racemase was engineered in a similar fashion and produced 12.6g l −1L -alanine from 1.8%glucose (Hols et al.1999).An Escherichia coli aceF ldhA double mutant containing pTrc99A-alaD plasmid produced 32g l −1racemic alanine in 27h during a two-stage (aerobic and anaerobic)fermentation with a yield of 0.63g alanine g −1glucose (Lee et al.2004).With further gene deletions and process optimization,the racemic alanine titer wasincreasedFig.1Alanine pathway in recombinant E.coli .a Native and recom-binant fermentation pathways.The foreign gene,G.stearothermophilus alaD ,is shown in bold .G.stearothermophilus alaD coding region and transcriptional terminator were integrated into the native ldhA gene under transcriptional control of the ldhA promoter.Solid stars represent deletions of native genes in XZ132.Note that the native biosynthetic route for alanine production is omitted for simplicity.ackA Acetate kinase,adhE alcohol/aldehyde dehydrogenase,alaD alanine dehydro-genase (Geobacillus stearothermophilus XL-65-6),aldA aldehyde dehydrogenase A,aldB aldehyde dehydrogenase B,alr alanine race-mase 1,dadX alanine racemase 2,frd fumarate reductase,gloA glyoxalase I,gloB glyoxalase II,gloC glyoxalase III,ldhA D -lactate dehydrogenase,mdh malate dehydrogenase,mgsA methylglyoxal synthase,pflB pyruvate –formate lyase,ppc phosphoenolpyruvate carboxylase,pta phosphate acetyltransferase.b Coupling of ATP production and growth to NADH oxidation and L -alanine production.Glucose is metabolized to pyruvate,ATP,and NADH.Energy conserved in ATP is utilized for growth and homeostasis,regenerating ADP.NADH is oxidized by alanine formation allowing glycolysis and ATP production to continueT a b l e 1C o m p a r i s o n o f a l a n i n e -p r o d u c i n g s t r a i n sO r g a n i s m s M o d i f i e d p r o p e r t yM e d i a ,s u b s t r a t e a n d p r o c e s s c o n d i t i o n sT i m e (h )A l a n i n e (g l −1)Y i e l d (%)L -A l a n i n ep u r i t y (%)R e f e r e n c e E .c o l i X Z 132I n t e g r a t e d G .s t e a r o t h e r m o p h i l u s a l a D ;Δp f l ,Δa c k A ,Δa d h E ,Δl d h A ,Δm g s A ,Δd a d XM i n e r a l m e d i u m ,b a t c h ,g l u c o s e 120g l −148.0114.095>99T h i s s t u d yA r t h r o b a c t e r o x y d a n s H A P -1M i n e r a l m e d i u m ,t w o -s t a g e f e d -b a t c h ,g l u c o s e 150g l −1120825560.0H a s h i m o t o a n d K a t s u m a t a 1998A .o x y d a n s D A N 75A l a n i n e r a c e m a c e d e f i c i e n tM i n e r a l m e d i u m ,t w o -s t a g e f e d -b a t c h ,g l u c o s e 150g l −1,0.2g l −1D -a l a n i n e120775198H a s h i m o t o a n d K a t s u m a t a 1998E c o l i A L 1(p O B P 1)P l a s m i d w i t h A .o x y d a n s H A P -1a l a D M i n e r a l m e d i u m ,g l u c o s e 20g l −1,l i m i t e d o x y g e n40841N o t r e p o r t e dK a t s u m a t a a n d H a s h i m o t o 1996C o r y n e b a c t e r i u m g l u t a m i c u m A L 107(p O B P 107)P l a s m i d w i t h A .o x y d a n s H A P -1a l a DC o r n s t e e p l i q u o r ,g l u c o s e 200g l −1,4g l −1D L -a l a n i n e ,l i m i t e d o x y g e n 707136>99K a t s u m a t a a n d H a s h i m o t o 1996Z y m o m o n a s m o b i l i s C P 4(p Z Y 73)P l a s m i d w i t h B .s p h a e r i c u s I F O 3525a l a D M i n e r a l s a l t s m e d i u m ,s i m p l e b a t c h ,g l u c o s e 50g l −126816N o t r e p o r t e dU h l e n b u s c h e t a l .1991L a c t o c o c c u s l a c t i s N Z 3950(p N Z 2650)P l a s m i d w i t h B .s p h a e r i c u s I F O 3525a l a D Δl d h AR i c h m e d i u m (M 17),g l u c o s e 18g l −117137085–90H o l s e t a l .1999L .l a c t i s P H 3950(p N Z 2650)P l a s m i d w i t h B .s p h a e r i c u s I F O 3525a l a D Δl d h A ,Δa l rR i c h m e d i u m (M 17),g l u c o s e 18g l −1,0.2g l −1D -a l a n i n e 17N o t k n o w nN o t k n o w n >99H o l s e t a l .1999E .c o l i A L S 887(p T r c 99A -a l a D )P l a s m i d w i t h B .s p h a e r i c u s I F O 3525a l a D Δl d h A ,Δa c e FY e a s t e x t r a c t ,t w o -s t a g e b a t c h ,g l u c o s e 50g l −1,a e r o b i c a i r 1l m i n −1273263N o t r e p o r t e d L e e e t a l .2004E .c o l i A L S 929(p T r c 99A -a l a D )P l a s m i d w i t h B .s p h a e r i c u s I F O 3525a l a D Δp f l ,Δp p s ,Δp o x B ,Δl d h A ,Δa c e E FY e a s t e x t r a c t a n d c a s a m i n o a c i d s ,t w o -s t a g e b a t c h (a e r o b i c c e l l g r o w t h a n d a n a e r o b i c f e r m e n t a t i o n )223486N o t r e p o r t e d S m i t h e t a l .2006E .c o l i A L S 929(p T r c 99A -a l a D )P l a s m i d w i t h B .s p h a e r i c u s I F O 3525a l a D Δp f l ,Δp p s ,Δp o x B ,Δl d h A ,Δa c e E FY e a s t e x t r a c t a n d c a s a m i n o a c i d s ,t w o -s t a g e f e d -b a t c h (a e r o b i c c e l l g r o w t h a n d a n a e r o b i c f e r m e n t a t i o n )4888100N o t r e p o r t e d S m i t h e t a l .2006to88g l−1in a more complex process with yields ap-proaching the theoretical maximum(Smith et al.2006). However,this strain produced only racemic alanine,utilized multicopy plasmids requiring antibiotic selection,and required complex media with a complex multistage fermen-tation process(Smith et al.2006).In this study,we developed novel biocatalysts that pro-duce chirally pure L-alanine in batch fermentations without using plasmid-containing biocatalysts,antibiotics,or com-plex nutrients.The resulting strains are based on a deriva-tive of E.coli W(strain SZ194)that produces D-lactate (Zhou et al.2006b).The ldhA gene in SZ194was replaced with a single,chromosomally integrated copy of the ALD gene from the thermophile,Geobacillus stearothermophilus XL-65-6(formerly B.stearothermophilus;Lai and Ingram 1993).After additional deletions of alanine racemase (dadX)and methylglyoxal synthase(mgsA)and metabolic evolution,the resulting strain produced L-alanine at high titers(over1M)and yields in batch fermentations using the mineral salts medium.Materials and methodsStrains,plasmids,media,and growth conditionsThe strains and plasmids used in this study are listed in Table2.Strain SZ194was previously engineered from a derivative of E.coli W(ATCC9637)and served as a starting point for constructions(Zhou et al.2006b).G. stearothermophilus XL-65-6(Lai and Ingram1993)was used for cloning the ALD gene.During sequencing of chro-mosomal genes,we discovered a20-year-old error in culture labeling.Strain SZ194,the parent used to construct the alanine strains,is a derivative of E.coli W(ATCC9637). Other constructs for ethanol production and lactate produc-tion that have been reported previously as derivatives of E. coli B are now known to be derivates of E.coli W(ATCC 9637).Primers used in this study are listed in Table3.During strain construction,cultures were grown aerobi-cally at30,37,or39°C in Luria broth(10g l−1Difco tryptone,5g l−1Difco yeast extract,and5g l−1NaCl) containing2%(w/v)glucose or5%(w/v)arabinose. Ampicillin(50mg l−1),tetracycline(12.5mg l−1), kanamycin(50mg l−1),or chloramphenicol(40mg l−1) were added as needed.For initial tests of fermentative alanine production,strains were grown without antibiotics at37°C in NBS mineral salts medium(Causey et al.2004) supplemented with100mM ammonia sulfate,1mM betaine,and2%(w/v)glucose.Fermentation experiments (2–12%sugar)were carried out in NBS medium and AM1 medium(Martinez et al.2007).Broth was maintained at pH 7by the automatic addition of5M NH4OH.Genetic methodsStandard methods were used for genomic deoxyribonucleic acid(DNA)extraction(Qiagen,Valencia,CA),polymerase chain reaction(PCR)amplification(Stratagene,La Jolla CA,and Invitrogen,Carlsbad,CA),transformation,plas-mid extration(Qiagen),and restriction endonuclease diges-tion(New England Biolabs,Ipswich,MA).Methods for foreign gene(alaD)integration and for chromosomal gene (mgsA and dadX)deletion are described below.DNA sequencing was provided by the University of Florida Interdisciplinary Center for Biotechnology Research.The Biocyc and Metacyc databases(Karp et al.2005)were instrumental in the design and completion of these studies. Cloning the alanine dehydrogenase gene alaD from G. stearothermophilus XL-65-6and detection of the enzyme activityThe primers for amplifying alaD from G.stearothermophilus XL-65-6were designed based on the alaD sequence of G. stearothermophilus strain10.The forward primers(5′–3′GGAAAAA GGAGGAAAAAGTG ATGAAGATCGG CATT)included the ribosomal-binding region(bold)and the amino terminus(italicized).The reverse primer(5′–3′GAA GGAGTTGATCATTGTTTAACGAGAGAGG)was down-stream from the putative transcriptional terminator region (Table3).ALD was verified in clones using an activity stain (Kuroda et al.1990).E.coli TOP10F′harboring plasmids containing alaD was grown on Luria–Bertani(LB)plates at 37°C,then transferred to a Whatman7.0-cm filter paper. The filter was immersed in10mM potassium phosphate buffer(pH7.2)and incubated for20min at80°C for lysis of the cells and denaturation of the E.coli proteins.The dried filter paper was assayed in a reaction mixture containing50mM L-alanine,50mM Tris–HCl buffer (pH9.0),0.625mM NAD+,0.064mM phenazine metho-sulfate,and0.24mM nitro blue tetrazolium.The cells with ALD appeared as blue spots on the filter.Integration of alaD into E.coli SZ194The alaD gene was integrated into the chromosomal ldhA gene of SZ194.The fragment(Sma I–Kpn I,1.7kb)con-taining a tet gene flanked by two FRT sites was isolated from pLOI2065and cloned into pLOI4211between a unique Bam HI site(Klenow-treated)and Kpn I site to produce plasmid pLOI4213(6.0kb).In this plasmid,transcription of alaD and tet are oriented in the same direction.The Apa I(treated with T4DNA polymerase to produce a blunt end)–Kpn I fragment(2.2kb)containing alaD and tet was isolated from pLOI4213and cloned into pLOI2395Table2 E.coli strains and plasmids used in this studyRelevant characteristics Source or referenceStrainsSZ194plfB frd adhE ackA deletions Zhou et al.2006bXZ103-110SZ194,ldhA::FRT-tet-FRT::This studyG.stearothermophilus alaDXZ111XZ105,ldhA::G.stearothermophilus alaD This studyXZ112XZ111,metabolic evolution in NBS medium with2%glucose This studyXZ113XZ112,metabolic evolution in NBS medium with5%glucose This studyXZ115XZ113,metabolic evolution in NBS medium with8%glucose This studyXZ121XZ115,mgsA deletion This studyXZ123XZ121,metabolic evolution in NBS medium with8%glucose This studyXZ126XZ123,dadX deletion This studyXZ129XZ126,metabolic evolution in NBS medium with8%glucose This studyXZ130XZ129,metabolic evolution in AM1medium with8%glucose This studyXZ131XZ130,metabolic evolution in AM1medium with10%glucose This studyXZ132XZ131,metabolic evolution in AM1medium with12%glucose This studyPlasmidspCR2.1-TOPO bla kan;TOPO TA cloning vector InvitrogenDatsenko and Wanner2000pKD46Blaγβexo(Red recombinase),temperature conditionalpSC101repliconpFT-A Bla flp,temperature conditional pSC101replicon Posfai et al.1997pEL04cat-sacB targeting cassette Lee et al.2001;Thomason et al.2005 pLOI2224kan;R6K conditional integration vector Martinez-Morales et al.1999pLOI2065bla;FRT-tet-FRT cassette Zhou et al.2003bpLOI2395bla;ldhA franked by two Asc I site Zhou et al.2003apLOI3421 1.8kbp SmaI fragment containing aac Wood et al.2005pLOI4151bla cat;cat-sacB cassette This studyalaD integrationThis studypLOI4211bla kan alaD;alaD(PCR)from G.stearothermophilus XL-65-6cloned into pCR2.1-TOPO vectorpLOI4213bla kan;alaD-FRT-tet-FRT Kpn I-Sma I fragment(FRT-tet-FRT)This studyfrom pLOI2065cloned into Kpn I-BamH I(blunted)site of pLOI4211This studypLOI4214bla kan;ldhA’-alaD-FRT-tet-FRT-ldhA”Apa I(blunted)-Kpn I fragment(alaD-FRT-tet-FRT)from pLOI4213cloned into ldhA at Hinc II-Kpn Isites of pLOI2395This studypLOI4215kan;ldhA’-alaD-FRT-tet-FRT-ldhA”Asc I fragment(ldhA’-alaD-FRT-tet-FRT-‘ldhA)from pLOI4214cloned into Asc I sites of pLOI2224mgsA deletionThis studypLOI4228bla kan;yccT’-mgsA-helD’(PCR)from E.coli W clonedinto PCR2.1-TOPO vectorThis studypLOI4229cat-sacB cassette PCR amplified from pLOI4151(Eco RV digested)cloned into mgsA in pLOI4228This studypLOI4230PCR fragment amplified from pLOI4228(using mgsA-1/mgsA-2primers),kinase treated,and self-ligateddadX deletionThis studypLOI4216bla kan;dadA’-dadX-cvrA’(PCR)from E.coli W clonedinto PCR2.1-TOPO vectorpLOI4218cat-sacB cassette PCR amplified from pLOI4151(Eco RV digested)This studycloned into dadX in pLOI4216This studypLOI4220PCR fragment amplified from pLOI4216(using dadX-4/dadX-5primers),kinase treated,and self-ligated(Hinc II to Kpn I sites)to produce pLOI4214(6.5kb).In this plasmid,ldhA ,alaD ,and tet genes are transcribed in the same direction.The Asc I fragment (4.3kb)containing these three genes was isolated from pLOI4214and cloned into the R6K integration vector pLOI2224to produce pLOI4215(6.2kb).Plasmid pLOI4215contains resistance genes for both tetracycline and kanamycin (Fig.2).The Asc I fragment (4.3kb)containing ldhA ,alaD ,and tet genes was isolated from pLOI4215,further cut by Xmn I to eliminate any remaining uncut plasmid DNA,and electroporated into SZ194containing the Red recombinase plasmid pKD46(Datsenko and Wanner 2000).Integrants were selected for tetracycline resistance,confirmed by sensitivity to kanamycin and ampicillin and by PCR analysis using the primers of ldhA and its neighboring genes ydbH and hslJ (Table 3).Deletion of mgsA and dadX genesA modified method for deleting E.coli chromosomal genes was developed using two steps of homologous recom-bination (Thomason et al.2005).With this method,no antibiotic genes or scar sequences remain on the chromo-some after gene deletion.In the first recombination,part of the target gene was replaced by a DNA cassette containing a chloramphenicol resistance gene (cat )and levansucrase gene (sacB ).In the second recombination,the cat –sacBcassette was removed by selection for resistance to sucrose.Cells containing the sacB gene accumulate levan during incubation with sucrose and are killed.Surviving recombi-nants are highly enriched for loss of the cat –sacB cassette.A new cassette was constructed as a template to facilitate gene deletions.The cat –sacB region was amplified from pEL04(Lee et al.2001;Thomason et al.2005)by PCR using the JM catsacB up Nhe I and JM catsacB down Nhe I primers (Table 3),digested with Nhe I,and ligated into the corresponding site in pLOI3421to produced pLOI4151.The cat –sacB cassette was amplified by PCR using pLOI4151as a template with the cat -up2and sacB -down2primers (Eco RV site included in each primer),digested with Eco RV ,and used in subsequent ligations.The mgsA gene and neighboring 500-bp regions (yccT ′–mgsA –helD ′,1,435bp)were amplified using the mgsA -up and mgsA -down primers and cloned into the pCR 2.1-TOPO vector (Invitrogen)to produce plasmid pLOI4228.A 1,000-fold diluted plasmid preparation of this plasmid served as a template for inside-out amplification using the mgsA -1and mgsA -2primers (both within the mgsA gene and facing outward).The resulting 4,958-bp fragment containing the replicon was ligated to the Eco RV-digested cat –sacB cassette from pLOI4151to produce pLOI4229(Fig.3a).This 4,958-bp fragment was also used to construct a second plasmid,pLOI4230(Fig.3b),by phosphorylation and self-ligation.In pLOI4230,the central region of mgsA is deleted (yccT ′–mgsA ′–mgsA ″–helD ′).After digestion of pLOI4229and pLOI4230with Xmn I (within the vector),each served as a template for amplifica-tion using the mgsA -up and mgsA -down primers to produce linear DNA for integration step 1(yccT ′–mgsA ′–cat –sacB –mgsA ″–helD ′)and step II (yccT ′–mgsA ′–mgsA ″–helD ′),respectively.After electroporation of the step 1fragment into XZ115containing pKD46(Red recombinase)and 2h ofTable 3Primers used in this study Primers SequencealaD -forward GGAAAAAGGAGGAAAAAGTGATGAA GATCGGCATTalaD -reverse GAAGGAGTTGATCATTGTTTAACGA GAGAGGldhA -forward AGTACCTGCAACAGGTGAAC ldhA -reverse CAGGCGACGGAATACGTCAT ldhA -up (ydbH )CTGATAACGCAGTTGCTGGA ldhA -down (hslJ )TTCATTAAATCCGCCAGCTTJM catsacB up NheI TTAGCTAGCATGTGACGGAAGATC ACTTCGJM catsacB down NheI CCGCTAGCATCAAAGGGAAAACTGT CCATATcat -up2AGAGAGGATATCTGTGACGGAAGAT CACTTCGsacB -down2AGAGAGGATATCGAATTGATCCGGT GGATGACmgsA -up CAGCTCATCAACCAGGTCAA mgsA -down AAAAGCCGTCACGTTATTGG mgsA -1AGCGTTATCTCGCGGACCGT mgsA -2AAGTGCGAGTCGTCAGTTCC dadX -up AGGCTACTCGCTGACCATTC dadX -down GGTTGTCGGTGACCAGGTAG dadX -4TGGGCTATGAGTTGATGTGC dadX -5CTGTATCGGACGGGTCATCTFig.2Integration vector used for chromosomal insertion of G.stearothermophilus alaD into E.coli ldhA .Sequence encoding the N-terminal and C-terminal regions are designated ldhA ′and ldhA ″,respectivelyincubation at 30°C to allow expression and segregation,recombinants were selected for chloramphenicol (40mg l −1)and ampicillin (50mg l −1)resistance in Luria broth at 30°C (18h).Three clones were selected,grown in Luria broth containing ampicillin and 5%(w/v)arabinose (to induce expression of red recombinase),and prepared for electro-poration.After electroporation with the step 2fragment,cells were incubated at 30°C for 4h and then transferred into a 250-ml flask containing 100ml of modified LB (100mM 3-(N -morpholino)propanesulfonic acid [MOPS]buffer added and NaCl omitted)containing 10%sucrose.After overnight incubation (30°C),clones were selected on modified LB plates (no NaCl;100mM MOPS added)containing 6%sucrose (39°C,16h).Resulting clones were tested for loss of ampicillin and chloramphenicol resistance.Construction was confirmed by PCR using the mgsA-up/down primer set.A clone containing a deletion in the central region of mgsA was selected and designated XZ121.The dadX gene was deleted in a manner analogous to that used to delete the mgsA gene.Primers for dadX deletion are shown in Table 3,and the corresponding plasmids are shown in Table 2.FermentationNBS mineral salts medium (Causey et al.2004)with 1mM betaine (Zhou et al.2006a )was used in the initial fermentation (pH 7.0).Preinoculum was grown by inocu-lating three colonies into a 250ml flask (100ml NBS medium,2%glucose,and 100mM ammonium sulfate).After 16h (37°C,120rpm),this preinoculum was diluted into 500-ml fermentation fleakers containing 300ml NBS medium (2–8%glucose,100mM ammonium sulfate,and 1mM betaine)with 33mg cell dry weight (CDW)l −1.In early experiments,pH was maintained at 7.0by automat-ically adding 2M potassium hydroxide.In later experi-ments,5M ammonium hydroxide was used to maintain pH,and a low salt medium,AM1(Martinez et al.2007),was used to replace the NBS medium for fermentation (8–12%glucose).AM1medium contains much less salt and has been optimized for E.coli .Metabolic evolutionCells from pH-controlled fermentations were serially transferred at 24-h intervals to facilitate metabolic evolution through competitive,growth-based selection (Fig.1b).At the beginning,sequentially transferred cultures were inoc-ulated with an initial density of 33mg CDW l −1.As growth increased,the inoculum was changed to a 1:100dilution and subsequently to a 1:300dilution.Periodically,clones were isolated from these experiments,assigned new strain designations,and frozen for storage.AnalysesCell mass was estimated by measuring the optical density at anic acids and glucose concentrations were mea-sured by high-performance liquid chromatography (HPLC,Underwood et al.2002).Analysis of fermentation products by mass spectroscopy and amino acid analyzer were provided by the University of Florida Interdisciplinary Center for Bio-technology Research.Alanine was found to be the predominant product.The alanine concentration and isomeric purity were further measured by HPLC using the Chiralpak MA(+)chiral column (Chiral Technologies,West Chester,PA).ResultCloning of the alanine dehydrogenase geneALD is found in Bacillus (and Geobacillus )species where it plays a pivotal role in energy generation during sporulation (Ohashima and Soda 1979;Kuroda et al.1990).ALD from B.sphaericus IFO3525has beenwidelyFig.3Plasmids used to delete mgsA .Plasmid pLOI4229(a )was used to delete the mgsA gene and insert the cat-sacB cassette in the first recombina-tion step.Plasmid pLOI4230(b )was used to remove the cat-sacB cassette to create a deletion devoid of foreign sequence.Se-quence encoding the N-terminal and C-terminal regions are des-ignated mgsA ′and mgsA ″,respectivelyused with varying degrees of success to engineer alanine production in recombinant bacteria(Uhlenbusch et al. 1991;Hols et al.1999;Lee et al.2004;Smith et al.2006). Selection of the B.sphaericus IFO3525is presumed to be due in part to the high specific activity(Ohashima and Soda 1979).In contrast,we have selected a thermostable ALD from the thermophile,G.stearothermophilus XL-65-6, based on our prior experience in expressing genes from this organism in recombinant E.coli(Burchhardt and Ingram 1992;Lai and Ingram1993;Lai and Ingram1995).The ribosomal-binding region,coding region,and tran-scriptional terminator of alaD were amplified from G. stearothermophilus XL-65-6and sequenced(EF154460in GenBank).The deduced amino acid sequence was identical to that reported for Geobacillus kaustophilus HTA426and very similar to G.stearothermophilus strain10(99%iden-tity)and G.stearothermophilus strain IFO12550(94% identity).The nucleotide sequence(65%identity)and the deduced ALD amino acid sequence(74%identity)were quite different from the B.sphaericus IFO3525gene,the gene pre-viously used for alanine production in recombinant bacteria.Modification of E.coli W for homoalanine productionE.coli W strain SZ194(pflB frdBC adhE ackA)was previously constructed to produce only D-lactic acid.All major fermentation pathways except lactate have been blocked in this strain by gene deletions(Fig.1a).To convert this strain to the production of alanine,part of the native ldhA-coding region was replaced by a DNA fragment containing the ribosomal-binding region,coding region,and transcriptional terminator of alaD from G. stearothermophilus XL-65-6.The promoterless alaD was oriented in the same direction as ldhA to allow expression from the native ldhA promoter(Fig.2).After electroporation,approximately500colonies were recovered with tetracycline resistance and sensitivity to kana-mycin,consistent with a double-crossover event.These colo-nies were further examined by PCR using ldhA forward and reverse primer set(Table3).Only eight colonies of the500 tested were correct based on an analysis of PCR fragments. These eight colonies were further verified using primer sets for alaD,ldhA forward and alaD reverse,alaD forward and ldhA reverse,and ldhA outside primers(Table3)and de-signated XZ103,XZ104,XZ105,XZ106,XZ107,XZ108, XZ109,and XZ110,respectively.These eight strains were initially tested in15-ml screw-cap tubes containing NBS medium with2%glucose and100mM ammonium sulfate, which were filled to the brim.Strain XZ105appeared to grow faster than the other strains(37°C for48h)and was selected for further development.XZ105was transformed with pFT-A,which contains an inducible flippase(FLP)recombinase(Martinez-Morales et al.1999;Posfai et al.1997).The chromosomal FRT-flanked tet gene in XZ105was removed by inducing the FLP recombinase.After growing in39°C to eliminate the temperature-sensitive plasmid pFT-A,resulting strain was designated XZ111.Expression of G.stearothermophilus alaD in XZ111is transcriptionally regulated by the ldhA promoter,the same promoter that regulates the production of lactate dehydrogenase(dominant fermentation pathway) in native E.coli.pH-controlled batch fermentation for alanine production Alanine production by strain XZ111was tested in500-ml fermentation vessels containing300ml NBS medium, 20g l−1glucose,100mM ammonium sulfate,and1mM betaine.Broth pH was automatically controlled by adding 2N potassium hydroxide.After96h,181mM alanine was produced.The alanine yield from total glucose was 81%(g/g),and84%based on glucose that had been metabo-lized.The chiral purity of L-alanine was96.1%(Table4). Very low levels of other products(lactate,succinate,ace-tate,ethanol)were present,typically below1mM.This result demonstrated that the integrated G.stearothermophilus alaD gene as a single chromosomal copy under the control of the native ldhA promoter can provide sufficient levels of ALD to support E.coli growth from the production of alanine as the sole fermentation product.Metabolic evolution of strain XZ111Although XZ111could accumulate alanine as the primary product,incubation times were long,and volumetric productivity was limited.When using a high-glucose concentration(80g l−1),growth and alanine productivity were further reduced(Table4).In this strain,adenosine triphosphate(ATP)production and growth are tightly coupled to NADH oxidation and alanine production by ALD(Fig.1b).This coupling provided a basis for strain improvement by selecting for increased growth during serial cultivation,i.e.,metabolic evolution.Cells with increased growth because of spontaneous mutations will successively displace their parents while coselecting for increased alanine productivity.Serial transfers of XZ111were carried out at24-h intervals in NBS mineral salts medium with1mM betaine.Cultures were first transferred in the medium containing20g l−1 glucose,and the pH was controlled by automatically adding 2N potassium hydroxide.However,after ten transfers to strain XZ112,little improvement was observed(data not shown).Because ammonia is essential for alanine pro-duction,it was thought that ammonia may be limiting for fermentation.Two normals potassium hydroxide containing 1N ammonia carbonate and5N ammonia hydroxide alone。
2一氯烟酸
烟酸及其衍生物烟酸一．市场信息烟酸化学名称为3-吡啶甲酸，又名尼古丁酸，维生素PP，是一种重要的精细化工中间体。
该产品在19世纪60年代就被人们发现，并由尼古丁氧化合成制得。
但是由于在该产品的应用方面的研究没有取得较大进展，因而长期以来未有大的发展，直到20世纪30年代，由于其在治疗糙皮病方面显现出了特有的疗效，所以引起了人们的普遍重视，众多科研单位加入了该产品的研制开发行列。
经过近70年的不断开发，目前该产品已开发出多条成熟的生产工艺路线，并且其应用领域覆盖医药、食品、饲料添加剂以及化工助剂等多个行业，呈现出良好的发展态势。
人们生活水平的提高和动物饲料产品的需求增长促进了全球烟酸／烟酰胺的快速发展有数据显示．2007年全球吡啶总产量的35％以上都是用来满足烟酰胺／烟酸的生产消耗．预计2012年还将保持这个比例目前，全球烟酸／烟酰胺总生产能力近6万其中。
美国和欧洲的产能在2．85万～2．95万t／a．其余的超过3万a的产能主要分布在亚洲，主要生产国家为中国、印度和Et本规模和产量较大的企业为瑞士的龙沙、美国的凡特鲁斯、印度吉友联公司、日本有机合成药品丁业株式会社以及中国台湾长春石油化学股份有限公司。
其中龙沙公司几十年来一直是世界最大的烟酸和烟酰胺生产企业．目前其产能近3万t／a，约占世界总产能的5O％。
美国Vitachem公司和Nepera公司总计生产能力为3500吨，年，此外，比利时德固赛、Antuerpen公司、德国德固赛以及日本的有机药品公司都是烟酸生产规模较大的公司。
目前，全球每年烟酸的需求量在2万吨左右，而且随着该产品在食品以及饲料添加剂方面应用量的逐年加大，其市场需求一直保持以每年3％左右的增长速度。
美国是世界上消耗量最高的国家，其每年的消费量约占全世界总量的45％左右，由于其国内生产能力不足，每年都进口大量的烟酸，烟酸胺以满足需求。
而西欧则是世界上烟酸的主要生产地，每年都向世界各地出口大量的产品。
盐酸吡哆醇分子量
盐酸吡哆醇分子量概述盐酸吡哆醇（Pyridoxine hydrochloride）是一种重要的维生素B6衍生物，化学名称为4,5-bis(hydroxymethyl)-2-methylpyridin-3-ol hydrochloride。
它在药物和医学领域具有广泛的应用，被广泛用于治疗B6维生素缺乏症和其他相关疾病。
了解盐酸吡哆醇的分子量对于药物研究和治疗非常重要。
盐酸吡哆醇的分子量盐酸吡哆醇的分子式为C8H12ClNO3，它包含了8个碳原子、12个氢原子、1个氯原子和3个氧原子。
根据元素的相对原子质量，我们可以计算出盐酸吡哆醇的分子量。
首先，我们需要查找各个原子的相对原子质量： - 碳（C）的相对原子质量为12.01 - 氢（H）的相对原子质量为1.008 - 氯（Cl）的相对原子质量为35.45 - 氧（O）的相对原子质量为16.00接下来，我们将各个原子的相对原子质量相加，并加上氯化氢（HCl）的相对原子质量（即氯的原子质量），即可计算出盐酸吡哆醇的分子量：分子量 = 8 *12.01 + 12 * 1.008 + 35.45 + 3 * 16.00 = 169.18 g/mol所以，盐酸吡哆醇的分子量为169.18 g/mol。
盐酸吡哆醇的应用盐酸吡哆醇在医学领域具有重要的应用价值。
它主要通过补充体内维生素B6来改善一系列相关疾病的症状。
1. 治疗维生素B6缺乏症维生素B6是一种重要的水溶性维生素，对身体各个系统的正常运转起着重要作用。
维生素B6缺乏可能导致贫血、皮肤炎症、神经系统障碍等症状。
盐酸吡哆醇可以作为补充维生素B6的药物，有效改善维生素B6缺乏引起的症状。
2. 预防和治疗妊娠反应妊娠反应是一种常见的孕期症状，包括恶心、呕吐等不适感。
盐酸吡哆醇可以帮助调节妊娠期的激素水平，减轻或预防妊娠反应的发生。
3. 支持正常神经系统发育和功能维生素B6在神经系统的发育和功能中扮演重要角色。
盐酸吡哆醇的补充可以促进神经系统的正常发育和功能，对于预防和治疗与神经系统相关的疾病具有积极作用。
苯佐卡因摩尔质量
苯佐卡因摩尔质量
苯佐卡因是一种局部麻醉药，常用于手术中减轻疼痛和不适感。
它的化学式为C13H20N2O，摩尔质量为236.31g/mol。
苯佐卡因主要通过阻止神经信号的传递来产生麻醉效果。
它能够阻止神经元内部的钠离子通道，从而阻止钠离子进入神经元内部，进而阻止神经信号的传递。
这种阻止神经信号传递的效果能够使疼痛信号无法到达大脑，从而减轻了疼痛和不适感。
苯佐卡因的应用范围非常广泛。
它可以用于局部麻醉手术，如皮肤切割、针刺、皮下注射等。
此外，它还可以用于治疗神经痛、痛经、牙痛等疾病。
苯佐卡因的用药方式也有多种。
最常见的方式是皮下注射或局部喷洒。
另外，还可以通过静脉注射或全身麻醉的方式使用。
当然，使用苯佐卡因也有一些需要注意的事项。
首先，苯佐卡因不能用于过敏者或对该药物有反应的人群。
其次，使用苯佐卡因时需要注意药物的剂量和使用方法，以免产生过量或过度麻醉的后果。
此外，苯佐卡因还可能产生一些副作用，如头晕、恶心、呕吐等。
苯佐卡因是一种非常常用的局部麻醉药。
其摩尔质量为236.31g/mol，主要通过阻止神经信号传递来产生麻醉效果。
它可以用于局部麻醉手术、治疗神经痛等疾病。
但在使用时需要注意剂
量和使用方法，并注意可能产生的副作用。
戈那瑞林
化合物简介
基本信息
物化性质
基本信息
中文名称：戈那瑞林 中文别名：黄体激素释放激素 英文名称：gonadorelin 英文别名：Glp-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2;(pyroglutamic acid)-His-Trp-SerTyr-Gly-Leu-Arg-Pro-Gly-NH2;Gonadorelin;GONADERELIN;luteinizing hormone-releasing hormone isoform I; CAS号：-09-2 分子式：C55H75N17O13 分子量：1182. 结构式： 精确质量：1181. PSA：472.
适应症
1.用于诊断下丘脑-垂体-生殖腺功能障碍。 2.治疗闭经与促性腺激素分泌不足和多滤泡性卵巢引起的不孕症。 3.戈那瑞林或其同类物布舍瑞林、戈舍瑞林、亮丙瑞林、那法瑞林和曲普瑞林还可用于避孕、隐睾症、恶性 肿瘤（尤其前列腺癌）、延迟的和提前的青春期。 4.还可用于子宫内膜异位。 5.用于促排卵以治疗下丘脑性闭经所致不孕、原发性卵巢功能不足，特别是对氯米芬无效的患者。 6.还用于小儿隐睾症及雄激素过多、垂体肿瘤等。 。
氨基酸比值取本品约2mg，置10ml量瓶中，加6mol/L盐酸溶液使溶解并稀释至刻度，摇匀，量取1ml置2ml硬 质安瓶中，在-5℃减压封口，置110℃加热24小时,冷却，启封，将内容物移至蒸馏瓶中，在减压下蒸干，残留物 加0.02mol/L盐酸溶液1ml溶解，摇匀，作为供试品溶液；另取与供试品溶液浓度相应的各个氨基酸对照品混合溶 液，作为对照品溶液。精密量取上述两种溶液各20μl，分别注入氨基酸分析仪，记录色谱图，按外标法以峰面 积计算，即得。以L一组氨酸、L一谷氨酸、L一亮氨酸、L一脯氨酸、甘氨酸和L一精氨酸摩尔数之和的七分之一 值为1，供试品溶液中各个氨基酸相应的摩尔比应符合以下规定：L一丝氨酸为0.7—1.05，L一谷氨酸为0.9—l.