Differential response of orthologous l,l-diaminopimelate aminotransferases (DapL) to enzyme inhibitory antibiotic lead compounds

l,l-Diaminopimelate aminotransferase (DapL) is an enzyme required for the biosynthesis of meso-diaminopimelate (m-DAP) and l-lysine (Lys) in some bacteria and photosynthetic organisms. m-DAP and Lys are both involved in the synthesis of peptidoglycan (PG) and protein synthesis. DapL is found in specific eubacterial and archaeal lineages, in particular in several groups of pathogenic bacteria such as Leptospira interrogans (LiDapL), the soil/water bacterium Verrucomicrobium spinosum (VsDapL) and the alga Chlamydomonas reinhardtii (CrDapL). Here we present the first comprehensive inhibition study comparing the kinetic activity of DapL orthologs using previously active small molecule inhibitors formerly identified in a screen with the DapL of Arabidopsis thaliana (AtDapL), a flowering plant. Each inhibitor is derived from one of four classes with different central structural moieties: a hydrazide, a rhodanine, a barbiturate, or a thiobarbituate functionality. The results show that all five compounds tested were effective at inhibiting the DapL orthologs. LiDapL and AtDapL showed similar patterns of inhibition across the inhibitor series, whereas the VsDapL and CrDapL inhibition patterns were different from that of LiDapL and AtDapL. CrDapL was found to be insensitive to the hydrazide (IC₅₀ >200 μM). VsDapL was found to be the most sensitive to the barbiturate and thiobarbiturate containing inhibitors (IC₅₀ ∼5 μM). Taken together, the data shows that the homologs have differing sensitivities to the inhibitors with IC₅₀ values ranging from 4.7 to 250 μM. In an attempt to understand the basis for these differences the four enzymes were modeled based on the known structure of AtDapL. Overall, it was found that the enzyme active sites were conserved, although the second shell of residues close to the active site were not. We conclude from this that the altered binding patterns seen in the inhibition studies may be a consequence of the inhibitors forming additional interactions with residues proximal to the active site, or that the inhibitors may not act by binding to the active site. Compounds that are specific for DapL could be potential biocides (antibiotic, herbicide or algaecide) that are nontoxic to animals since animals do not contain the enzymes necessary for PG or Lys synthesis. This study provides important information to expand our current understanding of the structure/activity relationship of DapL and putative inhibitors that are potentially useful for the design and or discovery of novel biocides.     

二氧化氯对球形棕囊藻的抑制和杀灭作用

以海洋赤潮生物—球形棕囊藻汕头株(Phaeocystis globosa,ST strain)为材料,研究了ClO2对不同起始藻密度的棕囊藻的抑制和杀灭作用．结果表明,ClO2对球形棕囊藻有明显的抑制和杀灭作用．藻密度为2．35×10^9cells·L-1时,高于0．74×10-2mmol·L-1的ClO2对棕囊藻生长有一定的抑制作用。高于2．96×10-2mmol·L-1的ClO2对棕囊藻具有显著的杀灭作用．藻密度与ClO2浓度之间存在一定的剂量关系．藻密度为2．35×10^9、1．18×10^9、4．70×10^8、1．18×10^8 cells·L-1时。其96h的有效杀藻浓度分别为2．96×10^-2、2．22×10^-2、1．48×10^-2和0．59×10^-2mmol·L^-1．藻密度越高。杀灭单位藻细胞所需ClO2的浓度越低．ClO2作为杀藻剂在赤潮治理中具有很好的应用前景．     

海水中苯扎溴铵除藻剂的紫外可见分光光度法测定

Benzalkonium bromide is a high-efficiency algaecide. Its concentration in seawater was measured by the method of spectrophotometer. The results indicated that the deposition appeared if the concentration of benzalkonium bromide in seawater was more than 50mg- L- 1 and affected the detection of benzalkonium bromide. But, there was a good linear relationship between concentration and absorbanee when the concentration was lower than 50mg-L-1 (R2= 0. 9996). Therefore, spectrophotometer could be used to detect benzalkonium bromide in seawater.     

湛江球形棕囊藻赤潮除藻试验

2005年4月,在广东湛江港赤潮区采集球形棕囊藻(Phaeoecystisglobosa),在室内用新洁尔灭和异噻唑啉酮两种除藻剂进行棕囊藻的杀灭试验。结果表明,新洁尔灭对赤潮水体中球形棕囊藻的72hLC50为0.96mgL-1;与新洁尔灭相比,异噻唑啉酮的除藻效果更为明显,其对球形棕囊藻的72hLC50为0.54mgL-1。新洁尔灭和异噻唑啉酮复配时具有协同作用,可提高除藻能力,新洁尔灭和异噻唑啉酮浓度比为1.3:0.3(配比为4.3:1)时除藻效果最优,2d的除藻率达到89%,协同指数为0.78。两种除藻剂对鲻鱼(Mugilcephalus)苗72h的毒性试验表明,新洁尔灭和异噻唑啉酮浓度分别在2.0和0.3mgL-1以下除藻是安全的。     

碘伏和异噻唑啉酮对球形棕囊藻去除的研究

研究了碘伏和异噻唑啉酮除藻剂对球形棕囊藻赤潮生物的灭杀和控制作用,结果表明,单独使用时,碘伏的最低有效浓度为30mg.L^-1,异噻唑啉酮最低有效浓度为0.30mg.L^-1,当两者复配时有协同作用,可提高它们的杀藻能力,碘伏与异噻唑啉酮浓度比为1.0:0.15时除藻效果最佳。     

特丁基三嗪对两种赤潮藻去除的实验研究

研究了特丁基三嗪(tertbutyl triazille)对塔玛亚历山大藻和球形棕囊藻两种赤潮生物的杀灭和控制作用。结果表明,特丁基三嗪能有效地控制和杀灭塔玛亚历山大藻和球形棕囊藻,其中96h杀灭塔玛亚历山大藻的有效浓度为0．2mgL^-1,杀灭球形棕囊藻的有效浓度为0.3mgL^-1。特丁基三嗪具有高效、作用时效长的特点,可能是一种比较理想的赤潮藻去除剂。