Pullulanase: Role in Starch Hydrolysis and Potential Industrial Applications

The use of pullulanase (EC 3.2.1.41) has recently been the subject of increased applications in starch-based industries especially those aimed for glucose production. Pullulanase, an important debranching enzyme, has been widely utilised to hydrolyse the α-1,6 glucosidic linkages in starch, amylopectin, pullulan, and related oligosaccharides, which enables a complete and efficient conversion of the branched polysaccharides into small fermentable sugars during saccharification process. The industrial manufacturing of glucose involves two successive enzymatic steps: liquefaction, carried out after gelatinisation by the action of α-amylase; saccharification, which results in further transformation of maltodextrins into glucose. During saccharification process, pullulanase has been used to increase the final glucose concentration with reduced amount of glucoamylase. Therefore, the reversion reaction that involves resynthesis of saccharides from glucose molecules is prevented. To date, five groups of pullulanase enzymes have been reported, that is, (i) pullulanase type I, (ii) amylopullulanase, (iii) neopullulanase, (iv) isopullulanase, and (v) pullulan hydrolase type III. The current paper extensively reviews each category of pullulanase, properties of pullulanase, merits of applying pullulanase during starch bioprocessing, current genetic engineering works related to pullulanase genes, and possible industrial applications of pullulanase.

[1]  J. Shaw,et al.  Production of High-maltose Syrup and High-protein Flour from Rice by an Enzymatic Method. , 1992, Bioscience, biotechnology, and biochemistry.

[2]  G. Haki,et al.  Developments in industrially important thermostable enzymes: a review. , 2003, Bioresource technology.

[3]  Saroj P. Mathupala,et al.  Characterization of an endo-Acting Amylopullulanase from Thermoanaerobacter Strain B6A , 1990, Applied and environmental microbiology.

[4]  A. Pugsley,et al.  Molecular characterization of pulA and its product, pullulanase, a secreted enzyme of Klebsielia pneumoniae UNF5023 , 1990, Molecular microbiology.

[5]  Reinder Johannes Leemhuis.  What makes cyclodextrin glycosyltransferase a transglycosylase , 2003 .

[6]  G. Antranikian,et al.  Characterization of amylolytic and pullulytic enzymes from thermophilic archaea and from a newFervidobacterium species , 1994, Applied Microbiology and Biotechnology.

[7]  W. M. Fogarty,et al.  Recent Advances in Microbial Amylases , 1990 .

[8]  R. Singhal,et al.  Induction of pullulanase production in Bacillus cereus FDTA-13. , 2007, Bioresource technology.

[9]  T. Imanaka,et al.  Purification and Characterization of Thermostable Pullulanase from Bacillus stearothermophilus and Molecular Cloning and Expression of the Gene in Bacillus subtilis , 1988, Applied and environmental microbiology.

[10]  Lubbert Dijkhuizen,et al.  Properties and applications of starch-converting enzymes of the α-amylase family , 2002 .

[11]  M. Rossi,et al.  Glycosyl hydrolases from hyperthermophiles , 1997, Extremophiles.

[12]  J. Rendleman.  Enhancement of cyclodextrin production through use of debranching enzymes , 1997, Biotechnology and applied biochemistry.

[13]  C. d’Enfert,et al.  Characterization and expression of the structural gene for pullulanase, a maltose-inducible secreted protein of Klebsiella pneumoniae , 1985, Journal of bacteriology.

[14]  Serge Pérez,et al.  The molecular structures of starch components and their contribution to the architecture of starch granules: A comprehensive review , 2010 .

[15]  G. Antranikian,et al.  Cloning, expression and biochemical characterisation of a unique thermostable pullulan-hydrolysing enzyme from the hyperthermophilic archaeon Thermococcus aggregans. , 2000, FEMS microbiology letters.

[16]  Yuzuru Suzuki,et al.  A hyperthermostable pullulanase produced by an extreme thermophile, Bacillus flavocaldarius KP 1228, and evidence for the proline theory of increasing protein thermostability , 1991, Applied Microbiology and Biotechnology.

[17]  R. Tharanathan,et al.  Grain legumes—a boon to human nutrition , 2003 .

[18]  P. Nigam,et al.  Enzyme and microbial systems involved in starch processing. , 1995 .

[19]  Ming-chu Chen,et al.  Structure and expression of an amylopullulanase gene from Bacillus stearothermophilus TS‐23 , 2001, Biotechnology and applied biochemistry.

[20]  Ajay Singh,et al.  Developments in the use of Bacillus species for industrial production. , 2004, Canadian journal of microbiology.

[21]  S. Radosta,et al.  Study of a preparative-scale process for the production of amylose , 2002 .

[22]  G. Antranikian,et al.  Purification and properties of a thermostable pullulanase from Clostridium thermosulfurogenes EM1 which hydrolyses both α-1,6 and α-1,4-glycosidic linkages , 1990, Applied Microbiology and Biotechnology.

[23]  J. Ko,et al.  Purification and biochemical characterization of pullulanase type I from Thermus caldophilus GK-24. , 1996, FEMS microbiology letters.

[24]  P. Johnsen.  Factors influencing the flavor quality of farm-raised catfish , 1989 .

[25]  R. E. Hebeda,et al.  CHAPTER 12 – Starches, Sugars, and Syrups , 1993 .

[26]  Suren Singh,et al.  Improved high thermal stability of pullulanase from a newly isolated thermophilic Bacillus sp. AN-7 , 2006 .

[27]  A. Salyers,et al.  Characterization of a neopullulanase and an alpha-glucosidase from Bacteroides thetaiotaomicron 95-1 , 1991, Journal of bacteriology.

[28]  A. Ariff,et al.  Direct fermentation of gelatinized sago starch to acetone–butanol–ethanol by Clostridium acetobutylicum , 2001 .

[29]  Stephen H. Brown,et al.  Characterization of Amylolytic Enzyme Activities Associated with the Hyperthermophilic Archaebacterium Pyrococcus furiosus , 1990, Applied and environmental microbiology.

[30]  Martyn Poliakoff,et al.  Sustainable technology: Green chemistry , 2007, Nature.

[31]  S. Ito,et al.  Purification and Some Properties of an Alkaline Pullulanase from Alkalophilic Bacillus sp.KSM-1876 , 1992 .

[32]  S. Jonjić,et al.  Modulation of natural killer cell activity by viruses. , 2010, Current opinion in microbiology.

[33]  W. Liebl,et al.  Isolation and analysis of genes for amylolytic enzymes of the hyperthermophilic bacterium Thermotoga maritima. , 1998, FEMS microbiology letters.

[34]  M. J. Yebra,et al.  Characterization of novel neopullulanase fromBacillus polymyxa , 1997 .

[35]  P. Bergquist,et al.  Cloning and sequence of a type I pullulanase from an extremely thermophilic anaerobic bacterium, Caldicellulosiruptor saccharolyticus. , 1997, Biochimica et biophysica acta.

[36]  C. Biliaderis,et al.  Amylolytic enzymes and products derived from starch: a review. , 1995, Critical reviews in food science and nutrition.

[37]  Teodoro Espinosa-Solares,et al.  Macroscopic mass and energy balance of a pilot plant anaerobic bioreactor operated under thermophilic conditions , 2006, Applied biochemistry and biotechnology.

[38]  W. Liebl,et al.  Cloning, Sequencing, and Characterization of a Heat- and Alkali-Stable Type I Pullulanase from Anaerobranca gottschalkii , 2004, Applied and Environmental Microbiology.

[39]  Y. Sakano,et al.  A classification of dextran-hydrolysing enzymes based on amino-acid-sequence similarities. , 1997, The Biochemical journal.

[40]  G. Antranikian,et al.  Starch-hydrolyzing enzymes from thermophilic archaea and bacteria. , 2002, Current opinion in chemical biology.

[41]  E. Farina,et al.  Degradation of dental plaque glucans and prevention of glucan formation using commercial enzymes , 2002 .

[42]  J. Robyt,et al.  Enzyme modification of starch granules: formation and retention of cyclomaltodextrins inside starch granules by reaction of cyclomaltodextrin glucanosyltransferase with solid granules. , 2000, Carbohydrate research.

[43]  C. Martin,et al.  Starch biosynthesis. , 1995, The Plant cell.

[44]  S. Bejar,et al.  Isolation and purification of an acidic pullulanase type II from newly isolated Bacillus sp. US149 , 2003 .

[45]  T. Imanaka,et al.  New type of pullulanase from Bacillus stearothermophilus and molecular cloning and expression of the gene in Bacillus subtilis , 1988, Journal of bacteriology.

[46]  D. Summers,et al.  Recombinant protein secretion in Escherichia coli. , 2005, Biotechnology advances.

[47]  Wolfgang Aehle,et al.  Enzymes in industry : production and applications , 2004 .

[48]  Kubo,et al.  The starch-debranching enzymes isoamylase and pullulanase are both involved in amylopectin biosynthesis in rice endosperm , 1999, Plant physiology.

[49]  B. Norman.  A Novel Debranching Enzyme for Application in the Glucose Syrup Industry , 1982 .

[50]  日本農芸化学会.  Agricultural and biological chemistry , 1961 .

[51]  Helmut. Uhlig,et al.  Industrial enzymes and their applications , 1998 .

[52]  M Takagi,et al.  Cloning of the aapT gene and characterization of its product, alpha-amylase-pullulanase (AapT), from thermophilic and alkaliphilic Bacillus sp. strain XAL601 , 1994, Applied and environmental microbiology.

[53]  G. Antranikian,et al.  Pullulanase Type I from Fervidobacterium pennavorans Ven5: Cloning, Sequencing, and Expression of the Gene and Biochemical Characterization of the Recombinant Enzyme , 1999, Applied and Environmental Microbiology.

[54]  T. Imanaka,et al.  Pattern of action of Bacillus stearothermophilus neopullulanase on pullulan , 1989, Journal of bacteriology.

[55]  K. Ozaki,et al.  Nucleotide and deduced amino acid sequences of an alkaline pullulanase from the alkaliphilic bacterium Bacillus sp. KSM-1876. , 2001, Biochimica et biophysica acta.

[56]  E. Schneider,et al.  Gene cloning, nucleotide sequence and biochemical properties of a cytoplasmic cyclomaltodextrinase (neopullulanase) from Alicyclobacillus acidocaldarius, reclassification of a group of enzymes. , 2000, FEMS microbiology letters.

[57]  J. Karkalas,et al.  Starch-composition, fine structure and architecture , 2004 .

[58]  S. Ito,et al.  Purification and characterization of an alkaline amylopullulanase with both α-1,4 and α-1,6 hydrolytic activity from alkalophilic Bacillus sp. KSM-1378 , 1995 .

[59]  C. R. Soccol,et al.  Advances in microbial amylases. , 2000, Biotechnology and applied biochemistry.

[60]  G. Antranikian,et al.  Isolation and characterization of a heat-stable pullulanase from the hyperthermophilic archaeon Pyrococcus woesei after cloning and expression of its gene in Escherichia coli , 1995, Applied and environmental microbiology.

[61]  P. Debeire,et al.  Purification and properties of the catalytic domain of the thermostable pullulanase type II from Thermococcus hydrothermalis , 2001, Biotechnology Letters.

[62]  R. Ohba,et al.  Purification, Crystallization and Some Properties of Intracellular Pullulanase from Aerobacter aerogenes , 1973 .

[63]  S. Bejar,et al.  Thermostable pullulanase type I from new isolated Bacillus thermoleovorans US105: cloning, sequencing and expression of the gene in E. coli , 2002 .

[64]  Yasunori Nakamura.  Some properties of starch debranching enzymes and their possible role in amylopectin biosynthesis , 1996 .

[65]  G. Antranikian,et al.  A New Thermoactive Pullulanase from Desulfurococcus mucosus: Cloning, Sequencing, Purification, and Characterization of the Recombinant Enzyme after Expression in Bacillus subtilis , 2000, Journal of bacteriology.

[66]  J. Shetty,et al.  Commodity scale production of sugars from starches. , 1999, Current opinion in microbiology.

[67]  K. Horikoshi.  Alkaliphiles: Some Applications of Their Products for Biotechnology , 1999, Microbiology and Molecular Biology Reviews.

[68]  B. Ranjbar,et al.  Purification and characterization of a novel amylopullulanase that converts pullulan to glucose, maltose, and maltotriose and starch to glucose and maltose , 2010 .

[69]  G. Antranikian,et al.  Extremophiles as a source of novel enzymes for industrial application , 1999, Applied Microbiology and Biotechnology.

[70]  Š. Janeček,et al.  Thermophilic archaeal amylolytic enzymes , 2000 .

[71]  K. Kawakami,et al.  Kinetics of condensation of glucose into maltose and isomaltose in hydrolysis of starch by glucoamylase , 1985, Biotechnology and bioengineering.

[72]  C. Mitchinson,et al.  Enzymes involved in the processing of starch to sugars , 1997 .

[73]  G. Antranikian,et al.  Purification and Properties of a Thermostable Pullulanase from a Newly Isolated Thermophilic Anaerobic Bacterium, Fervidobacterium pennavorans Ven5 , 1997, Applied and environmental microbiology.

[74]  Stephen H. Brown,et al.  Characterization of Amylolytic Enzymes, Having Both α-1,4 and α-1,6 Hydrolytic Activity, from the Thermophilic Archaea Pyrococcus furiosus and Thermococcus litoralis , 1993 .

[75]  W. Steiner,et al.  Highly thermostable amylase and pullulanase of the extreme thermophilic eubacterium Rhodothermus marinus: production and partial characterization. , 2003, Bioresource technology.

[76]  D. Topping,et al.  Starches, resistant starches, the gut microflora and human health. , 2000, Current issues in intestinal microbiology.

[77]  S. Jobling.  Improving starch for food and industrial applications. , 2004, Current opinion in plant biology.