L-Leucine-p-nitroanilide | CAS#4178-93-2 | substrate

MedKoo Cat#: 597478 | Name: L-Leucine-p-nitroanilide

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Description:

WARNING: This product is for research use only, not for human or veterinary use.

L-Leucine-p-nitroanilide is used to detect/evaluate functionality of amino peptidases. The compound is used in an ELISA assay to detect biological functionality of chicken amino peptidase produced by E. coli. It is also employed to characterize the native IRAP (placental leucine amino peptidase). Leu-pNA, Chromogenic Substrate is documented for use with pure samples as well as cell extracts in vitro.

Chemical Structure

L-Leucine-p-nitroanilide

CAS#4178-93-2

Theoretical Analysis

MedKoo Cat#: 597478

Name: L-Leucine-p-nitroanilide

CAS#: 4178-93-2

Chemical Formula: C12H17N3O3

Exact Mass: 251.1270

Molecular Weight: 251.29

Elemental Analysis: C, 57.36; H, 6.82; N, 16.72; O, 19.10

Price and Availability

Size	Price	Availability
500mg	USD 250.00	2 weeks
1g	USD 400.00	2 weeks
2g	USD 650.00	2 Weeks
5g	USD 950.00	2 weeks

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Synonym

L-Leucine-p-nitroanilide; H-Leu-pna

IUPAC/Chemical Name

(S)-2-amino-4-methyl-N-(4-nitrophenyl)pentanamide

InChi Key

AXZJHDNQDSVIDR-NSHDSACASA-N

InChi Code

InChI=1S/C12H17N3O3/c1-8(2)7-11(13)12(16)14-9-3-5-10(6-4-9)15(17)18/h3-6,8,11H,7,13H2,1-2H3,(H,14,16)/t11-/m0/s1

SMILES Code

CC(C)C[C@H](N)C(NC1=CC=C([N+]([O-])=O)C=C1)=O

Appearance

Solid powder

Purity

>98% (or refer to the Certificate of Analysis)

Shipping Condition

Shipped under ambient temperature as non-hazardous chemical. This product is stable enough for a few weeks during ordinary shipping and time spent in Customs.

Storage Condition

Dry, dark and at 0 - 4 C for short term (days to weeks) or -20 C for long term (months to years).

Solubility

Soluble in DMSO

Shelf Life

>3 years if stored properly

Drug Formulation

This drug may be formulated in DMSO

Stock Solution Storage

0 - 4 C for short term (days to weeks), or -20 C for long term (months).

HS Tariff Code

2934.99.03.00

More Info

Product Data

Product Data

Certificate of Analysis

View CoA: current batch, Lot#A22F07A22

Safety Data Sheet (SDS)

View Safety Data Sheet (SDS)

Instruction

View handling instruction

Biological target:

L-Leucine-p-nitroanilide is used to detect/evaluate functionality of amino peptidases.

In vitro activity:

To screen lactic acid bacteria for starter cultures in cheese production, 21 Lactococcus strains previously isolated from natural fermented milk and koumiss made by herdsman families in the Xinjiang, Gansu, and Qinghai provinces of China were evaluated for optimal growth temperature, acidification activity, proteolytic activity, aminopeptidase activity, and autolytic activity. For aminopeptidase activity, strains X9C2 and T1C2 displayed the highest activities of 30.56 and 27.70 U/mg of protein using L-leucine-p-nitroanilide as substrate, respectively. Reference: J Dairy Sci. 2011 Apr;94(4):1691-6. https://pubmed.ncbi.nlm.nih.gov/21426956/

In vivo activity:

TBD

Preparing Stock Solutions

The following data is based on the product molecular weight 251.29 Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.

Concentration / Solvent Volume / Mass	1 mg	5 mg	10 mg
1 mM	1.15 mL	5.76 mL	11.51 mL
5 mM	0.23 mL	1.15 mL	2.3 mL
10 mM	0.12 mL	0.58 mL	1.15 mL
50 mM	0.02 mL	0.12 mL	0.23 mL

Formulation protocol:

1. Ma CL, Zhang LW, Yi HX, Du M, Han X, Zhang LL, Feng Z, Zhang YC, Li Q. Technological characterization of lactococci isolated from traditional Chinese fermented milks. J Dairy Sci. 2011 Apr;94(4):1691-6. doi: 10.3168/jds.2010-3738. PMID: 21426956. 2. Premarathne AA, Leung DW. Characterization of activity of a potential food-grade leucine aminopeptidase from kiwifruit. Enzyme Res. 2010 Nov 4;2010:517283. doi: 10.4061/2010/517283. PMID: 21076540; PMCID: PMC2976069.

In vitro protocol:

In vivo protocol:

TBD

1: Lin SH, Van Wart HE. Effect of cryosolvents and subzero temperatures on the hydrolysis of L-leucine-p-nitroanilide by porcine kidney leucine aminopeptidase. Biochemistry. 1982 Oct 26;21(22):5528-33. PubMed PMID: 7171573. 2: Shimasue A, Sugimoto Y, Tsubokura T. [No blank method using L-leucine-p-nitroanilide as substrate for the assay of serum leucine aminopeptidase (author's transl)]. Rinsho Byori. 1977 Feb;25(2):157-60. Japanese. PubMed PMID: 558393. 3: Zhu X, Barman A, Ozbil M, Zhang T, Li S, Prabhakar R. Mechanism of peptide hydrolysis by co-catalytic metal centers containing leucine aminopeptidase enzyme: a DFT approach. J Biol Inorg Chem. 2012 Feb;17(2):209-22. doi: 10.1007/s00775-011-0843-2. Epub 2011 Sep 15. PubMed PMID: 21918843. 4: Hernández-Moreno AV, Villaseñor F, Medina-Rivero E, Pérez NO, Flores-Ortiz LF, Saab-Rincón G, Luna-Bárcenas G. Kinetics and conformational stability studies of recombinant leucine aminopeptidase. Int J Biol Macromol. 2014 Mar;64:306-12. doi: 10.1016/j.ijbiomac.2013.12.023. Epub 2013 Dec 22. PubMed PMID: 24368112. 5: Landau R, Laverrière A, Bischof P, Irion O, Morales M, Cohen M. Alteration of circulating Placental Leucine Aminopeptidase (P-LAP) activity in preeclampsia. Neuro Endocrinol Lett. 2010;31(1):63-6. PubMed PMID: 20150869. 6: Wang H, Xu W, Cao J, Wang W. Rapid screening of aminopeptidase N inhibitors by capillary electrophoresis with electrophoretically mediated microanalysis. Electrophoresis. 2015 Jan;36(2):319-25. doi: 10.1002/elps.201400283. Epub 2014 Dec 17. PubMed PMID: 25348092. 7: Premarathne AA, Leung DW. Characterization of activity of a potential food-grade leucine aminopeptidase from kiwifruit. Enzyme Res. 2010 Nov 4;2010:517283. doi: 10.4061/2010/517283. PubMed PMID: 21076540; PubMed Central PMCID: PMC2976069. 8: Hegazi FZ, Abo-Elnaga IG. Proteolytic activity of crude cell-free extract of Lactobacillus casei and Lactobacillus plantarum. Nahrung. 1987;31(3):225-32. PubMed PMID: 3112575. 9: Brecher AS, Moehlman TA, Hann W. Utilization of chymotrypsin as a sole carbon and (or) nitrogen source by Escherichia coli. Can J Microbiol. 1992 Apr;38(4):290-5. PubMed PMID: 1611554. 10: Watanabe T, Matsuura M, Seto K. Proteolytic activity of mycoplasmas and ureaplasmas isolated freshly from human saliva. Med Microbiol Immunol. 1985;173(5):251-5. PubMed PMID: 3881657. 11: Marinova M, Dolashki A, Altenberend F, Stevanovic S, Voelter W, Tchorbanov B. Purification and characterization of L-phenylalanine aminopeptidase from chick-pea cotyledons (Cicer arietinum L.). Protein Pept Lett. 2009;16(2):207-12. PubMed PMID: 19200046. 12: Fialho MC, Moreira NR, Zanuncio JC, Ribeiro AF, Terra WR, Serrão JE. Prey digestion in the midgut of the predatory bug Podisus nigrispinus (Hemiptera: Pentatomidae). J Insect Physiol. 2012 Jun;58(6):850-6. doi: 10.1016/j.jinsphys.2012.03.009. Epub 2012 Mar 20. PubMed PMID: 22440738. 13: Nishimura C, Suzuki H, Tanaka N, Yamaguchi H. Bleomycin hydrolase is a unique thiol aminopeptidase. Biochem Biophys Res Commun. 1989 Sep 15;163(2):788-96. PubMed PMID: 2506856. 14: Ma CL, Zhang LW, Yi HX, Du M, Han X, Zhang LL, Feng Z, Zhang YC, Li Q. Technological characterization of lactococci isolated from traditional Chinese fermented milks. J Dairy Sci. 2011 Apr;94(4):1691-6. doi: 10.3168/jds.2010-3738. PubMed PMID: 21426956. 15: Gao JJ, Xue X, Gao ZH, Cui SX, Cheng YN, Xu WF, Tang W, Qu XJ. LYP, a bestatin dimethylaminoethyl ester, inhibited cancer angiogenesis both in vitro and in vivo. Microvasc Res. 2011 Sep;82(2):122-30. doi: 10.1016/j.mvr.2011.05.008. Epub 2011 Jun 12. Review. PubMed PMID: 21664364. 16: Vander Jagt DL, Baack BR, Hunsaker LA. Purification and characterization of an aminopeptidase from Plasmodium falciparum. Mol Biochem Parasitol. 1984 Jan;10(1):45-54. PubMed PMID: 6363922. 17: Gao JJ, Gao ZH, Zhao CR, Yuan Y, Cui SX, Zhang XF, Cheng YN, Xu WF, Tang W, Qu XJ. LYP, a novel bestatin derivative, inhibits cell growth and suppresses APN/CD13 activity in human ovarian carcinoma cells more potently than bestatin. Invest New Drugs. 2011 Aug;29(4):574-82. doi: 10.1007/s10637-010-9391-9. Epub 2010 Jan 29. PubMed PMID: 20111888. 18: Doi H, Kawakami M. Purification of aminopeptidase from Australian classified barley flour. Int J Biochem Cell Biol. 1997 Feb;29(2):345-52. PubMed PMID: 9147136. 19: Frey ST, Guilmet SL, Egan RG 3rd, Bennett A, Soltau SR, Holz RC. Immobilization of the aminopeptidase from Aeromonas proteolytica on Mg2+/Al3+ layered double hydroxide particles. ACS Appl Mater Interfaces. 2010 Oct;2(10):2828-32. doi: 10.1021/am1005095. PubMed PMID: 20825230; PubMed Central PMCID: PMC2981069. 20: Cui SX, Qu XJ, Gao ZH, Zhang YS, Zhang XF, Zhao CR, Xu WF, Li QB, Han JX. Targeting aminopeptidase N (APN/CD13) with cyclic-imide peptidomimetics derivative CIP-13F inhibits the growth of human ovarian carcinoma cells. Cancer Lett. 2010 Jun 28;292(2):153-62. doi: 10.1016/j.canlet.2009.11.021. Epub 2009 Dec 29. PubMed PMID: 20042271.