Biomaterial Database

Purification and characterization of a papaya (Carica papaya L.) pectin methylesterase isolated from a commercial papain preparation. 2012

Prasanna Vasu, and Brett J Savary, and Randall G Cameron

Arkansas Biosciences Institute, Arkansas State University, Jonesboro, AR, USA.

We purified a Carica papaya pectin methylesterase (CpL-PME; EC 3.1.1.11) from a commercial papain preparation. This CpL-PME was separated from the abundant cysteine endopeptidases activities using sequential hydrophobic interaction and cation-exchange chromatographies and then purified by affinity chromatography using Sepharose-immobilized kiwi PME inhibitor protein to obtain a single electrophoretically homogeneous protein. The enzyme was purified 92-fold with 38% yield, providing a specific activity of 1200 U/mg. The molecular weight was determined to be 35,135 by MALDI-TOF-MS in linear mode. MALDI-TOF-MS peptide mass fingerprinting following trypsin digestion indicated CpL-PME represents a novel Carica PME isoform. The CpL-PME required salt for activity, and it showed a broad activity range (pH 6-9) and moderate thermostability (optimum ca. 70°C). A calcium-insensitive methylated lime pectin treated with CpL-PME to reduce degree of methylesterification by 6% converted the substrate to high calcium sensitivity, indicating a processive mode of action. These properties support further research to apply CpL-PME to tailor pectin nanostructure.

UI	MeSH Term	Description	Entries
D010206	Papain	A proteolytic enzyme obtained from Carica papaya. It is also the name used for a purified mixture of papain and CHYMOPAPAIN that is used as a topical enzymatic debriding agent. EC 3.4.22.2.	Tromasin
D010368	Pectins	High molecular weight polysaccharides present in the cell walls of all plants. Pectins cement cell walls together. They are used as emulsifiers and stabilizers in the food industry. They have been tried for a variety of therapeutic uses including as antidiarrheals, where they are now generally considered ineffective, and in the treatment of hypercholesterolemia.	Calcium Pectinate,Methoxy Pectin,Methoxylpectin,Methoxypectin,Pectin,Pectinic Acid,Zinc Pectinate,Pectin, Methoxy,Pectinate, Calcium,Pectinate, Zinc
D002845	Chromatography	Techniques used to separate mixtures of substances based on differences in the relative affinities of the substances for mobile and stationary phases. A mobile phase (fluid or gas) passes through a column containing a stationary phase of porous solid or liquid coated on a solid support. Usage is both analytical for small amounts and preparative for bulk amounts.	Chromatographies
D005638	Fruit	The fleshy or dry ripened ovary of a plant, enclosing the seed or seeds.	Berries,Legume Pod,Plant Aril,Plant Capsule,Aril, Plant,Arils, Plant,Berry,Capsule, Plant,Capsules, Plant,Fruits,Legume Pods,Plant Arils,Plant Capsules,Pod, Legume,Pods, Legume
D013058	Mass Spectrometry	An analytical method used in determining the identity of a chemical based on its mass using mass analyzers/mass spectrometers.	Mass Spectroscopy,Spectrometry, Mass,Spectroscopy, Mass,Spectrum Analysis, Mass,Analysis, Mass Spectrum,Mass Spectrum Analysis,Analyses, Mass Spectrum,Mass Spectrum Analyses,Spectrum Analyses, Mass
D029441	Carica	A plant genus of the family Caricaceae, order Violales, subclass Dilleniidae, class Magnoliopsida. It is the source of edible fruit and PAPAIN.	Papaya,Carica papaya,Carica papayas,Caricas,Papayas,papaya, Carica