33605-72-0Relevant articles and documents
High isolated yields in thermolysin-catalysed synthesis of Z-L-aspartyl-L-phenylalanine methyl ester in toluene at controlled water activity
De Martin, Luigi,Ebert, Cynthia,Gardossi, Lucia,Linda, Paolo
, p. 3395 - 3397 (2001)
Z-L-Aspartyl-L-phenylalanine methyl and ethyl esters were synthesised enzymatically in toluene by means of the zinc protease thermolysin adsorbed onto Celite R-640, which is a porous support able to control the hydration of the protein. The conversion of the two derivatised amino acids into the desired products was complete, leading to >90% isolated yields. Moreover, working with equimolar concentrations of the reactants no purification steps were required. Thermolysin adsorbed onto Celite R-640 is shown to be a practical tool to synthesise biologically active peptides in organic media.
Synthesis of an Aspartame Precursor by Immobilized Thermolysin in an Organic Solvent
Oyama, Kiyotaka,Nishimura, Shigeaki,Nonaka, Yuji,Kihara, Kei-ichi,Hashimoto, Tsutomu
, p. 5241 - 5242 (1981)
The synthesis of N-(benzyloxycarbonyl)-L-aspartyl-L-phenylalanine methyl ester, the precursor of the synthetic sweetener aspartame, from N-(benzyloxycarbonyl)-L-aspartic acid an L-phenylalanine methyl ester was carried out in an apparent single phase of the organic solvent by using thermolysin immobilized with various methods.
Enzymatic peptide synthesis in organic solvent with different zeolites as immobilization matrixes
Xing, Guo-Wen,Li, Xuan-Wen,Tian, Gui-Ling,Ye, Yun-Hua
, p. 3517 - 3522 (2000)
A series of zeolite immobilized α-chymotrypsin and thermolysin with microporous Y zeolites (HY, NH4, NaY) and mesoporous dealuminized DAY zeolites (HDAY, HNH4DAY) as matrixes have been prepared to catalyze peptide bond formation in organic solvents for the first time. The results indicated that most zeolite immobilized enzymes were active for peptide syntheses in organic media, and still had catalytic activity to some extent after being reused five times. According to the results, the immobilization effect of microporous Y zeolite was better than that of mesoporous DAY zeolite, suggesting that microporous Y zeolite can form more powerful hydrogen bonds with enzyme molecules since there are more hydroxyl groups on the Y zeolite than on the DAY zeolite. In addition, the influences of some reaction conditions such as reaction time and water content of the solvent on the enzymatic peptide synthesis were also studied and optimized. For the two kinds of proteases, NH4Y zeolite did not show its advantages for thermolysin, but was more suitable for α-chymotrypsin as an immobilization matrix. 2000 Elsevier Science Ltd.
Synthesis of aspartame by thermolysin: An x-ray structural study
Birrane, Gabriel,Bhyravbhatla, Balaji,Navia, Manuel A.
, p. 706 - 710 (2014)
Protease mediated peptide synthesis (PMPS) was first described in the 1930s but remains underexploited today. In most PMPS, the reaction equilibrium is shifted toward synthesis by the aqueous insolubility of product generated. Substrates and proteases are selected by trial and error, yields are modest, and reaction times are slow. Once implemented, however, PMPS reactions can be simple, environmentally benign, and readily scalable to a commercial level. We examined the PMPS of a precursor of the artificial sweetener aspartame, a multiton peptide synthesis catalyzed by the enzyme thermolysin. X-ray structures of thermolysin in complex with aspartame substrates separately, and after PMPS in a crystal, rationalize the reaction's substrate preferences and reveal an unexpected form of substrate inhibition that explains its sluggishness. Structure guided optimization of this and other PMPS reactions could expand the economic viability of commercial peptides beyond current high-potency, low-volume therapeutics, with substantial green chemistry advantages.
Effects of Solvents and Additives on the Reaction of N-Benzyloxycarbonyl-L-Aspartic Anhydride with L-Phenylalanine Methyl Ester ( Synthesis of Aspartame)
Yang, Chin-Ping,Su, Chein-Shyong
, p. 5186 - 5191 (1986)
N-Benzyloxycarbonyl-L-aspartic anhydride (Z-L-aspartic anhydride) was reacted with L-phenylalanine methyl ester.The nature of the ring-opening reaction was effected by the organic solvent.In Me2SO, DMF, and dimethylacetamide the β-isomer of Z-L-aspartyl-L-phenylalanine methyl ester (ZAPM) was predominant while in glacial acetic acid, esters, ketones, ethers, chlorohydrocarbons, acetonitrile, toluene, etc. the α-isomer of ZAPM prevailed.Especially in glacial acetic acid, the yield of α-form ZAPM reached more than 85percent.The results of reactions in mixed solvents like acetonitrile-Me2SO and ethyl acetate-acetonitrile showed that the yield of α-ZAPM changed linearly with a change in the composition of the mixed two solvents.The reaction in acetonitrile-acetic acid and Me2SO-acetic acid showed that the addition of acetic acid not only caused a solvent effect by itself, but that the acid also affected the reaction.Therefore, the yield of α-ZAPM was increased by use of a mixed solvent containing only a small amount of glacial acetic acid.A mechanism to explain these experimental phenomena is presented.
Functional Capsule Membranes. Part 29. Thermolysin-immobilized Capsule Membranes as Bioreactors in the Synthesis of a Dipeptide (Precursor of Aspartame) in an Organic Solvent
Okahata, Yoshio,Ijiro, Kuniharu
, p. 91 - 96 (1988)
Thermolysin (TLN) was covalently immobilized onto a large, ultrathin nylon capsule membrane grafted with poly- using glutaraldehyde.When the TLN-immobilized capsule containing a buffer solution (pH 7) in the inner aqueous phase was immersed in a chloroform solution of N-benzyloxycarbonyl-L-aspartic acid (Z-L-Asp) and L-phenylalanine methyl ester (L-PheOMe) with shaking at 40 deg C, the dipeptide (Z-L-Asp-L-PheOMe) was produced efficiently in the outer chloroform solution.From the Lineweaver-Burk plot, condensation in the aqueous-organic solvent involves initial binding of Z-L-Asp to the enzyme to form the Z-L-Asp-enzyme complex and then attack by L-PheOMe on the complex as the rate-determining step to form the peptide linkage.The TLN-capsule system can be used repeatedly without denaturation of protein structures by organic solvents because the enzyme on the capsule membrane is protected by buffer solutions coming from the inside.The enzyme-immobilized capsule membrane is a new bioreactor in aqueous-organic heterophases.
One-pot syntheses of dissymmetric diamides based on the chemistry of cyclic monothioanhydrides. Scope and limitations and application to the synthesis of glycodipeptides
Crich, David,Sasaki, Kaname,Rahaman, Md Yeajur,Bowers, Albert A.
supporting information; experimental part, p. 3886 - 3893 (2009/10/20)
(Chemical Equation Presented) Opening cyclic monothioanhydrides by amines provides a convenient entry into amido thioacids that can be trapped in situ by 2,4-dinitrobenzenesulfonamides, by electron-deficient azides, or by amines in the presence of Sanger's reagent leading, in each case, to dissymmetric diamides in what can be considered a one-pot, three-component coupling sequence. The use of monothiomaleic anhydride and bifunctional nucleophiles such as amino thiols provides access to heterocyclic amides. The low reactivity of cyclic monothioanhydrides toward alcohols enables the use of methanol as solvent and obviates the need for the protection of alcohols in the various reaction components. Reaction of N-benzyloxycarbonyl-L-aspartic monothioanhydride with unprotected glycosyl amines, followed by capture of the thioacid intermediate with N-sulfonyl amino acid derivatives results in a three-component convergent synthesis of glycosylated peptides.
Biocatalytic properties of thermolysin immobilized on polyvinyl alcohol cryogel
Belyaeva,Smirnova,Lysogorskaya,Oksenoit,Timofeeva,Lozinskii,Filippova
, p. 435 - 441 (2008/12/21)
Preparations with different contents of thermolysin were obtained by the immobilization of the enzyme on granulated polyvinyl alcohol cryogel. Their activity and stability in an aqueous medium and in mixtures of polar organic solvents of different composition were investigated. The catalytic properties of the preparations in reactions of peptide bond formation were studied, and the optimal amount of the biocatalyst, the concentrations of initial reagents, and the ratios of organic solvents and water necessary for effective enzymatic peptide synthesis catalyzed by immobilized thermolysin were determined. A series of peptides of the general formula Z-Ala-Ala-Xaa-pNA, where Xaa = Leu, Ile, Phe, Val, or Ala, were synthesized, and the immobilized enzyme was shown to retain substrate specificity in an organic medium.
Control of hydrolysis and condensation activities of thermolysin by ultrasound irradiation
Kawasaki, Takayoshi,Hoshino, Yu,Ishizu, Yukari,Mizushiro, Yu,Okahata, Yoshio
, p. 1602 - 1603 (2007/10/03)
Hydrolysis and condensation reactions of peptides catalyzed by thermolysin can be reversibly controlled by on/off ultrasound irradiation depending on its frequency region. Copyright
Bromelain catalyzed synthesis of peptides in organic solvent
Tai, Dar-Fu,Fu, Shu-Lin
, p. 179 - 183 (2007/10/03)
For the first time, immobilized bromelain was shown to maintain high catalytic activity in organic solvent and to form peptide bonds. It requires only 7 hours to obtain Cbz-Gly-L-Leu-OMe in 85% yield. The precursor of aspartame (Cbz-L-Asp-L-Phe-OMe) and other dipeptides were also synthesized by this method.
