| In recent years,biocatalysis has been widely used in the pharmaceutical industry.Compared with the defects of traditional chemical methods,including harsh reaction conditions,the need for toxic transition metal catalysts and volatile organic compounds,primarily solvents,and insufficient stereoselectivity in a single step.As well as possessing excellent enantioselectivity,the ability of enzymes to operate in aqueous media and at ambient temperature and pH mean they are seen as a potential "green alternative" to metal-catalyzed reactions.With the continuous application of aminotransferases in industry,more and more attempts have been made to discover and screen more efficient and economical transaminases.However,transaminase catalysis also has some drawbacks,for example,traditional methods for screening transaminases generally have the disadvantages of long time consumption,high detection cost,and reaction equilibrium problem of transaminase during the catalytic process.Based on the above situation,this article constructed a rapid and efficient method for the screening of transaminases:Firstly,1-naphthylmethylamine was chosen as the amino donor and pyruvate was used as amino acceptor in transaminase reactions.After transaminase reaction,1-naphthylmethylamine was converted into 1-naphthaldehyde,which could then react with oxalyl dioxime by adding to the system to form a hydrazide compound,the hydrazide compound has strong fluorescence emission at a wavelength of 486 nm.Therefore,the activity of transaminase was qualitatively analyzed by fluorescence detection system at 486 nm.At the same time,another set of enzymatic reaction equilibrium elimination system was constructed in this paper.By using benzylamine as an amino donor,by-products after transaminase reaction were formed.Benzaldehyde can react with oxalyl dioxime,breaking the balance of the enzymatic reaction and prompting the transaminase reaction to obtain more desired amine.In addition,we have found that the hydrazide compounds synthesized in high-throughput screens are susceptible to coordination reactions of metal ions,causing significant changes in their optical properties,and the amount of metal ions can be quantified by the change in their fluorescence intensity.At the same time,aluminum,as one of the most abundant metal elements in the earth’s crust,is extensively used in modern society involving industrial fields and daily life.On the other hand,the potential toxicity of aluminum ion has attracted considerable attention.According to reports,high levels of aluminum ions in the human body can cause serious damage to the nervous system,thereby inducing various diseases.Therefore,how to quickly and efficiently detect the content of low-concentration aluminum ions in nature has become an issue that people are increasingly concerned about.In order to detect the content of Al3+,a simple fluorescent probe L(2-hydroxy-1-naphthaldehyde oxalyl dioxime)was designed and synthesized as a ligand.This probe can recognize Al3+ through fluorescence "switching" response.Compared to other metal ions,probe L has excellent selectivity for Al3+,excluding the interference of other metal ions with L fluorescence intensity.At the same time,this program found a linear relationship between the fluorescence intensity of L and the metal ion concentration,thus achieving the accurate detection of Al3+ content in the liquid phase. |
