Microbial Amidases and their Industrial Applications: A Review

Abstract

Among the pediatric cancer in developed countries, acute leukemia ¬constitutes the major part with affecting 30-45 per 1,000,000 children each year. The effect of treatment varies with differences in patients clinical, immunologic and genetic characteristics. Therefore, the search for efficient drugs to solve this problem is being continued worldwide. Although several kinds of treatments are available, enzyme therapy is equally effective. Enzymes have been used as drugs; likewise L-asparaginase and L-glutaminase had received much attention in recent years due to their anticarcinogenic potential. These enzymes constitute one of the most biotechnologically and biomedically important group of therapeutic enzymes accounting for about 40% of the total worldwide enzyme sales. Various sources are found to be good producers of the enzymes: bacteria, fungi along with some of the plant and animal species. Food and Drug Administration and World Health Organization have approved L-asparaginase for the effective treatment of acute lymphoblastic leukemia and lymphsarcoma. L-asparaginase and L-glutaminase break down L-asparagine or L-glutamine into L-aspartic acid or L-glutamic acid, respectively, and ammonia. L-asparagine depletion results in nutritional deprivation, inhibition of protein synthesis, and subsequent apoptotic cell death in lymphoblasts. On the other hand, the ability of L-asparaginase to selectively hydrolyzes L-asparagine into L-aspartateis a potential way to reduce the amount of free L-asparagine in the starting materials of food production, thus reducing the imminent risk of generating a potential carcinogenic and neurotoxic acrylamide that formed from L-asparagine and reducing sugars in carbohydrate-containing foods (such as snacks and biscuits) when they are heated above 120oC. Therefore, the present review is an attempt to compile information on the sources, antino plastic action and industrial application of microbial amidases enzymes.

A Rapid Method for the Selection of Amidohydrolases from Metagenomic Libraries by Applying Synthetic Nucleosides and a Uridine Auxotrophic Host

“…The hydrolytic activity of the recombinant enzymes was assayed qualitatively by testing 29 different substrates ( Figure 4A): N 4 -acylated (2'-deoxy)cytidines and cytosines (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16); N 2 -acetylisocytosines (17); chromogenic substrates such as p-nitroacetanilide (18) and p-nitrobenzanilide (19); various p-nitrophenyl (pNP) esters (20)(21)(22)(23)(24); terephthalate derivatives bis(2-hydroxyethyl) terephthalate (26), dimethyl terephthalate (27), and monomethyl terephthalate (28); and the beta-lactam nitrocefin (29). Hydrolysis of chromogenic substrates (18)(19)(20)(21)(22)(23)(24)(25) was analysed spectrophotometrically at 405 nm, and that of other substrates was analysed spectrophotometrically at 240-320 nm and using TLC or HPLC-MS ( Figures S4-S10). All the reactions were monitored up to 24 h. The 8 hydrolases selected from the metagenomic libraries showed different activity profiles towards N-acylated nucleosides harbouring short, bulky aliphatic, aromatic, and heterocyclic groups ( Figure 4B).…”

“…At present, amidases are used for the biosynthesis 2 of 12 of β-lactam antibiotics [13], and the production of (S)-4-fluorophenylglycine (a chiral intermediate for the synthesis of antiemetic drugs) [14], nicotinic [15] and 2-chloronicotinic acids [16], and the herbicide L-phosphinothricin [17]. L-asparaginase and L-glutaminase are used as oncolytic enzymes [18], and in the food industry for the production of acrylamide-free food (to reduce the formation of acrylamide during the frying of starchy foods at high temperatures) [18].…”

Studies on optimization of L-glutaminase production under submerged fermentation from marine Bacillus subtilis JK-79

“…Hence, the use of L-glutaminase deprives the tumor cells from L-glutamine and causes selective death of L-glutamine dependant tumor cells. Thus, it can act as a possible candidate for enzyme therapy (Hensley et al, 2013;El-Ghonemy, 2014). In recent years, L-glutaminase in combination with or as an alternative to L-asparginase could be used as in enzyme therapy for cancer particularly leukemia (Sabu, 2003).…”

Identification and Molecular Characterization of Genes Coding Pharmaceutically Important Enzymes from Halo-Thermo Tolerant Bacillus

“…L-glutaminase and L-asparaginase) are commonly used of therapeutic agents accounting for about 40% of the total worldwide enzyme sales. 45 Moreover, microbial glutaminases are enzymes with emerging potential in food industries. 44 One of the potential application of recombinant B. licheniformis glutaminase is the bioconversion of glutamine to flavorenhancing glutamic acid in fermented food products.…”

“…L-glutaminase such as L-asparaginase is very significant anticancer enzyme in the treatment of acute lymphoblastic leukemia and other kinds of cancer through the Lglutamine amino acid depletion in cancerous cells. 44,45 These enzymes (i.e. L-glutaminase and L-asparaginase) are commonly used of therapeutic agents accounting for about 40% of the total worldwide enzyme sales.…”