Pectinase

Last updated
Endopolygalacturonase I
Identifiers
Organism Aspergillus niger
SymbolpgaI
UniProt P26213
Other data
EC number 3.2.1.15
Search for
Structures Swiss-model
Domains InterPro

Pectinases are a group of enzymes that breaks down pectin, a polysaccharide found in plant cell walls, through hydrolysis, transelimination and deesterification reactions. [1] [2] Commonly referred to as pectic enzymes, they include pectolyase, pectozyme, and polygalacturonase, one of the most studied and widely used[ citation needed ] commercial pectinases. It is useful because pectin is the jelly-like matrix which helps cement plant cells together and in which other cell wall components, such as cellulose fibrils, are embedded. Therefore, pectinase enzymes are commonly used in processes involving the degradation of plant materials, such as speeding up the extraction of fruit juice from fruit, including apples and sapota. Pectinases have also been used in wine production since the 1960s. [3] The function of pectinase in brewing is twofold, first it helps break down the plant (typically fruit) material and so helps the extraction of flavors from the mash. Secondly the presence of pectin in finished wine causes a haze or slight cloudiness. Pectinase is used to break this down and so clear the wine.

Contents

Pectinases can be extracted from fungi such as Aspergillus niger . The fungus produces these enzymes to break down the middle lamella in plants so that it can extract nutrients from the plant tissues and insert fungal hyphae. If pectinase is boiled it is denatured (unfolded) making it harder to connect with the pectin at the active site, and produce as much juice.

Characterizations

Pectinase is a generic term used for a group of enzymes that catalyse the degradation of pectin by hydrolysis, trans-elimination, as well as de-esterification reactions. The degradation of pectic polymers is mainly caused by exo- and endo-polygalacturonases (exo- and endo-PGs), pectate and pectin lyases (PLs), pectin methylesterase (PME) and acetylesterase (PAE), β-galactosidase (β-Gal), and α-L-arabinofuranosidase (α-L-Af), among others. [4] [5]

The following table shows a summary of enzymes involved in pectin degradation. HG-PUL = homogalacturonan polysaccharide utilization loci; RG-I PUL = rhamnogalacturonan I polysaccharide utilization loci.

PULCAZyme familiesEC numberAccepted nameReaction
HG-PULPL1EC4.2.2.2Pectate lyaseEliminative cleavage of (1 → 4)-α-D-galacturonan to give oligosaccharides with 4-deoxy-α-D-galact-4-enuronosyl groups at their non-reducing ends
PL1EC4.2.2.10Pectin lyaseEliminative cleavage of (1 → 4)-α-D-galacturonan methyl ester to give oligosaccharides with 4-deoxy-6-O-methyl-α-D-galact-4-enuronosyl groups at their non-reducing ends
GH28EC3.2.1.67Galacturonan 1,4-α-galacturonidase[(1 → 4)-α-D-galacturonide]n + H2O = [(1 → 4)-α-D-galacturonide]n-1 + D-galacturonate
GH28EC3.2.1.82Exo-poly-α-digalacturonosidase[(1 → 4)-α-D-galacturonosyl]n + H2O = α-D-galacturonosyl-(1 → 4)-D-galacturonate + [(1 → 4)-α-D-galacturonosyl]n-2
GH28EC3.2.1.15Endo-polygalacturonase(1,4-α-D-galacturonosyl)n+m + H2O = (1,4-α-D-galacturonosyl)n + (1,4-α-D-galacturonosyl)m
CE8EC3.1.1.11PectinesterasePectin + n H2O = n methanol + pectate
CE4EC3.1.1.6AcetylesteraseAn acetic ester + H2O = an alcohol + acetate
RG-I PULPL9EC4.2.2.23Rhamnogalacturonan endolyaseEndotype eliminative cleavage of L-α-rhamnopyranosyl-(1 → 4)-α-D-galactopyranosyluronic acid bonds of rhamnogalacturonan I domains in ramified hairy regions of pectin leaving L-rhamnopyranose at the reducing end and 4-deoxy-4,5-unsaturated D-galactopyranosyluronic acid at the non-reducing end
GH28EC3.2.1.171Rhamnogalacturonan hydrolaseEndohydrolysis of α-D-GalA-(1 → 2)-α-L-Rha glycosidic bond in the rhamnogalacturonan I backbone with initial inversion of anomeric configuration releasing oligosaccharides with β-D-GalA at the reducing end.
GH2EC3.2.1.146β-GalactofuranosidaseHydrolysis of terminal non-reducing β-D-galactofuranosides, releasing galactose
GH138EC3.2.1.173Rhamnogalacturonan galacturonohydrolaseExohydrolysis of the α-D-GalA-(1 → 2)-α-L-Rha bond in rhamnogalacturonan oligosaccharides with initial inversion of configuration releasing D-galacturonic acid from the non-reducing end of rhamnogalacturonan oligosaccharides
GH105EC3.2.1.172Unsaturated rhamnogalacturonyl hydrolase2-O-(4-deoxy-β-L-threo-hex-4-enopyranuronosyl)-α-L-rhamnopyranose + H2O = 5-dehydro-4-deoxy-D-glucuronate + L-rhamnopyranose
GH106EC3.2.1.174Rhamnogalacturonan rhamnohydrolaseExohydrolysis of the α-L-Rha-(1 → 4)-α-D-GalA bond in rhamnogalacturonan oligosaccharides with initial inversion of configuration releasing β-L-rhamnose from the non-reducing end of rhamnogalacturonan oligosaccharides
GH43EC3.2.1.99Arabinan endo-1,5-α-L-arabinanaseEndohydrolysis of (1 → 5)-α-arabinofuranosidic linkages in (1 → 5)-arabinans
GH51EC3.2.1.55Non-reducing end α-L-arabinofuranosidaseHydrolysis of terminal non-reducing α-L-arabinofuranoside residues in α-L-arabinosides
GH146EC3.2.1.185Non-reducing end β-L-arabinofuranosidaseβ-L-arabinofuranosyl-(1 → 2)-β-L-arabinofuranose + H2O = 2 β-L-arabinofuranose
GH53EC3.2.1.89Arabinogalactan endo-β-1,4-galactanaseThe enzyme specifically hydrolyses (1 → 4)-β-D-galactosidic linkages in type I arabinogalactans
GH43EC3.2.1.145Galactan 1,3-β-galactosidaseHydrolysis of terminal, non-reducing β-D-galactose residues in (1 → 3)-β-D-galactopyranans
GH27EC3.2.1.88Non-reducing end β-L-arabinopyranosidaseRemoval of a terminal β-L-arabinopyranose residue from the non-reducing end of its substrate
GH43EC3.2.1.181Galactan endo-β-1,3-galactanaseThe enzyme specifically hydrolyses β-1,3-galactan and β-1,3-galactooligosaccharides
CE12EC3.1.1.86Rhamnogalacturonan acetylesteraseHydrolytic cleavage of 2-O-acetyl- or 3-O-acetyl groups of α-D-galacturonic acid in rhamnogalacturonan I.
RG-II PULGH43EC3.2.1.55Non-reducing end α-L-arabinofuranosidaseHydrolysis of terminal non-reducing α-L-arabinofuranoside residues in α-L-arabinosides
CE19EC3.1.1.11PectinesterasePectin + n H2O = n methanol + pectate
GH142EC3.2.1.185Non-reducing end β-L-arabinofuranosidaseβ-L-arabinofuranosyl-(1 → 2)-β-L-arabinofuranose + H2O = 2 β-L-arabinofuranose
GH78EC3.2.1.40α-L-RhamnosidaseHydrolysis of terminal non-reducing α-L-rhamnose residues in α-L-rhamnosides
GH33EC3.2.1.1243-deoxy-2-OctulosonidaseEndohydrolysis of the β-ketopyranosidic linkages of 3-deoxy-D-manno-2-octulosonate in capsular polysaccharides
GH95EC3.2.1.631,2-α-L-FucosidaseMethyl-2-α-L-fucopyranosyl-β-D-galactoside + H2O = L-fucose + methyl β-D-galactoside
GH2EC3.2.1.31β-Glucuronidasea β-D-glucuronoside + H2O = D-glucuronate + an alcohol
GH2EC3.2.1.23β-galactosidaseHydrolysis of terminal non-reducing β-D-galactose residues in β-D-galactosides
GH138EC3.2.1.173Rhamnogalacturonan galacturonohydrolaseExohydrolysis of the α-D-GalA-(1 → 2)-α-L-Rha bond in rhamnogalacturonan oligosaccharides with initial inversion of configuration releasing D-galacturonic acid from the non-reducing end of rhamnogalacturonan oligosaccharides
GH141EC3.2.1.51

Properties

Crystal structures

All pectinase enzyme structures include a prism-shaped right-handed cylinder made up of seven to nine parallel β-helices. The three parallel β-helices that create the prism shape of the structure are referred to as PB1, PB2 and PB3, with PB1 and PB2 creating an antiparallel β and PB3 sitting perpendicularly to PB2. All substrate binding sites of the various esterases, hydrolases, and lyases are located on an outer cleft of the central parallel β-helix structure between protruding loops on the structure and PB1. [6]

Optimum environment

As with all enzymes, pectinases have an optimum temperature and pH at which they are most active. For example, a commercial pectinase might typically be activated at 45 to 55 °C and work well at a pH of 3.0 to 6.5. [3]

Reaction pathway

Pectinases depolymerise pectin through hydrolysis, trans-elimination and deesterification reaction processes, breaking down the ester bond that holds together the carboxyl and methyl groups in pectin. [7]

Endo-polygalacturonase progresses through a reaction along the following pathway: [8]

(1,4-alpha-D-galacturonosyl)n+m + H2O = (1,4-alpha-D-galacturonosyl)n + (1,4-alpha-D-galacturonosyl)m

Occurrence and Application

Pectinase in nature

Pectinase enzymes used today are naturally produced by fungi and yeasts (50%), insects, bacteria and microbes (35%) and various plants (15%), [9] but cannot be synthesized by animal or human cells. [10] In plants, pectinase enzymes hydrolyze pectin that is found in the cell wall, allowing for new growth and changes to be made. The chemical and structural properties of pectin is especially prone to changes in the fruit due to solubilization and enzymatic degradation which are considered to be the key processes responsible for the softening of fruit during ripening. Structural changes that occur in the middle lamella and primary cell wall during ripening result in cell separation and softening of the tissues. The molecular components of primary walls are modified during fruit ripening by the temporally and spatially regulated action of endogenous enzymes. [11] [12]

Similarly to their role in plants, pectinases break down pectin during the developmental stage of fungi.

Industrial uses

Pectinase enzymes play various roles in both the fruit juice and wine industries. They are used for clarification in fruit juices and also speed up fruit juice extraction through enzymatic liquefaction of fruit pulp. In addition, pectinase enzymes aid in formation of pulpy products in the fruit juice industry. Pectinase enzymes are used for extracting juice from purée. This is done when the enzyme pectinase breaks down the substrate pectin and the juice is extracted. The enzyme pectinase lowers the activation energy needed for the juice to be produced and catalyzes the reaction.

Pectinases are useful in the wine industry by extracting anthocyanin from the fruit, effectively intensifying the wine coloring. [7] Pectinase can also be used to extract juices from cell walls of plants cells.

Pectinases are also used for retting in the textile industry. [13] Addition of chelating agents or pretreatment of the plant material with acid enhance the effect of the enzyme.

References

Creative Commons by small.svg  This article incorporates text by Luna Barrera-Chamorro, África Fernandez-Prior, Fernando Rivero-Pino and Sergio Montserrat-de la Paz available under the CC BY 4.0 license.

  1. Sakai T, Sakamoto T, Hallaert J, Vandamme EJ (1993). "Pectin, pectinase and protopectinase: production, properties, and applications". Advances in Applied Microbiology. 39: 213–94. doi:10.1016/s0065-2164(08)70597-5. PMID   8213306.
  2. Singh, Ram Sarup; Singh, Taranjeet; Pandey, Ashok (2019-01-01), Singh, Ram Sarup; Singhania, Reeta Rani; Pandey, Ashok; Larroche, Christian (eds.), "Chapter 1 - Microbial Enzymes—An Overview", Advances in Enzyme Technology, Biomass, Biofuels, Biochemicals, Elsevier, pp. 1–40, ISBN   978-0-444-64114-4 , retrieved 2021-10-20
  3. 1 2 "Pectinase". Enzyme India. Archived from the original on 26 March 2010. Retrieved 26 March 2010.
  4. 1 2 Satapathy, Sonali; Rout, Jyoti Ranjan; Kerry, Rout George; Thatoi, Hrudayanath; Sahoo, Santi Lata (2020-08-06). "Biochemical Prospects of Various Microbial Pectinase and Pectin: An Approachable Concept in Pharmaceutical Bioprocessing". Frontiers in Nutrition. 7 117. doi: 10.3389/fnut.2020.00117 . ISSN   2296-861X. PMC   7424017 . PMID   32850938.
  5. Barrera-Chamorro, Luna; Fernandez-Prior, África; Rivero-Pino, Fernando; Montserrat-de la Paz, Sergio (January 2025). "A comprehensive review on the functionality and biological relevance of pectin and the use in the food industry". Carbohydrate Polymers. 348 (Pt A) 122794. doi: 10.1016/j.carbpol.2024.122794 . PMID   39562070.
  6. Gummadi, Sathyanarayana N.; Manoj, N.; Kumar, D. Sunil (2007), Polaina, Julio; MacCabe, Andrew P. (eds.), "Structural and Biochemical Properties of Pectinases" , Industrial Enzymes: Structure, Function and Applications, Dordrecht: Springer Netherlands, pp. 99–115, doi:10.1007/1-4020-5377-0_7, ISBN   978-1-4020-5377-1 , retrieved 2021-10-20
  7. 1 2 Saranaj, P; Naidu, M.A. (2014). "Microbial Pectinases: A Review". ResearchGate.
  8. "BRENDA - Information on EC 3.2.1.15 - endo-polygalacturonase". brenda-enzymes.org. Retrieved 2021-10-20.
  9. Melton, Laurence (2019). Encyclopedia of Food Chemistry (Volume 2 ed.). Elsevier. p. 271.
  10. Saranaj, P; Naidu, M.A. (2014). "Microbial Pectinases: A Review". ResearchGate.
  11. Zheng, Ling; Xu, Yinxiao; Li, Qian; Zhu, Benwei (December 2021). "Pectinolytic lyases: a comprehensive review of sources, category, property, structure, and catalytic mechanism of pectate lyases and pectin lyases". Bioresources and Bioprocessing. 8 (1) 79. doi: 10.1186/s40643-021-00432-z . ISSN   2197-4365. PMC   10992409 . PMID   38650254.
  12. Liu, Dongjie; Zhou, Weiwei; Zhong, Yuming; Xie, Xi; Liu, Huifan; Huang, Hua; Wang, Qin; Xiao, Gengsheng (July 2023). "Involvement of branched RG-I pectin with hemicellulose in cell–cell adhesion of tomato during fruit softening" . Food Chemistry. 413 135574. doi:10.1016/j.foodchem.2023.135574. PMID   36739644.
  13. Rebello, Sharrel; Anju, Mohandas; Aneesh, Embalil Mathachan; Sindhu, Raveendran; Binod, Parameswaran; Pandey, Ashok (13 July 2017). "Recent advancement sin the production and application of microbial pectinases: an overview" (PDF). Reviews in Environmental Science and Bio/Technology. 16 (3): 381–394. Bibcode:2017RESBT..16..381R. doi:10.1007/s11157-017-9437-y. S2CID   90607593.