TITLE

Enhanced levan production using chitin-binding domain fused levansucrase immobilized on chitin beads

AUTHOR(S)

Chiang, Chung-Jen; Wang, Jen-You; Chen, Po-Ting; Chao, Yun-Peng

PUB. DATE

March 2009

SOURCE

Applied Microbiology & Biotechnology;Mar2009, Vol. 82 Issue 3, p445

SOURCE TYPE

Academic Journal

DOC. TYPE

Article

ABSTRACT

Levan is a homopolymer of fructose which can be produced by the transfructosylation reaction of levansucrase (EC 2.4.1.10) from sucrose. In particular, levan synthesized by Zymomonas mobilis has found a wide and potential application in the food and pharmaceutical industry. In this study, the immobilization of Z. mobilis levansucrae (encoded by levU) was attempted for repeated production of levan. By fusion levU with the chitin-binding domain (ChBD), the hybrid protein was overproduced in a soluble form in Escherichia coli. After direct absorption of the protein mixture from E. coli onto chitin beads, levansucrase tagged with ChBD was found to specifically attach to the affinity matrix. Subsequent analysis indicated that the linkage between the enzyme and chitin beads was substantially stable. Furthermore, with 20% sucrose, the production of levan was enhanced by 60% to reach 83 g/l using the immobilized levansucrase as compared to that by the free counterpart. This production yield accounts for 41.5% conversion yield (g/g) on the basis of sucrose. After all, a total production of levan with 480 g/l was obtained by recycling of the immobilized enzyme for seven times. It is apparent that this approach offers a promising way for levan production by Z. mobilis levansucrase immobilized on chitin beads.

ACCESSION #

36520069

 

Related Articles

• The intracellular sucrase (SacA) of Zymomonas mobilis is not involved in sucrose assimilation. Aít-Abdelkader, Nadra; De Caro, Alain; Guzzo, Jean; Michel, Gérard P. F.; Baratti, Jacques C. // Biotechnology Letters;Mar2000, Vol. 22 Issue 6, p461 

  The intracellular sucrase SacA from Zymomonas mobilis was purified to homogeneity from a recombinant E. coli strain containing the SacA gene under an expression system. The protein was monomeric with a molecular mass of 58 kDa. The sucrase activity was maximal at 25 °C and thermal stability...

• Serine substitution for cysteine residues in levansucrase selectively abolishes levan forming activity. Senthilkumar, Velusamy; Busby, Stephen J.W.; Gunasekaran, Paramasamy // Biotechnology Letters;Oct2003, Vol. 25 Issue 19, p1653 

  Levansucrase is responsible for levan formation during sucrose fermentation of Zymomonas mobilis, and this decreases the efficiency of ethanol production. As thiol modifying agents decrease levan formation, a role for cysteine residues in levansucrase activity has been examined using derivatives...

• Improvement of P450BM-3 whole-cell biocatalysis by integrating heterologous cofactor regeneration combining glucose facilitator and dehydrogenase in E. coli. Schewe, Hendrik; Kaup, Bjoern-Arne; Schrader, Jens // Applied Microbiology & Biotechnology;Feb2008, Vol. 78 Issue 1, p55 

  Escherichia coli BL21, expressing a quintuple mutant of P450BM-3, oxyfunctionalizes α-pinene in an NADPH-dependent reaction to α-pinene oxide, verbenol, and myrtenol. We optimized the whole-cell biocatalyst by integrating a recombinant intracellular NADPH regeneration system through...

• Levan production using mutant strains of Zymomonas mobilis in different culture conditions. Muro, Ana Carolina; Rodríguez, Emilio; Abate, Carlos M.; Siñeriz, Faustino // Biotechnology Letters;Oct2000, Vol. 22 Issue 20, p1639 

  Several levan hyperproducing mutants of Zymomonas mobilis strains were selected by mutagenesis with N-methyl-N-nitro-nitrosoguanidine and caffeine. Highest levan production (41 g l-1) was obtained with a mutant strain HL 29 in a culture medium containing 200 g sucrose l-1 and 0.5 g (NH4)2SO4 l-1...

• Purification of food-grade oligosaccharides using immobilised cells of Zymomonas mobilis. Crittenden, R. G.; Playne, M. J. // Applied Microbiology & Biotechnology;Mar2002, Vol. 58 Issue 3, p297 

  Immobilised cells of the bacterium Zymomonas mobilis were used to remove glucose, fructose, and sucrose from food-grade oligosaccharide mixtures. Unpurified fructo-, malto-, isomalto-, gentio-, and inulin-oligosaccharides, containing total carbohydrate concentrations of 300 g l–1, were...

• Comparison of characteristics of levan produced by different preparations of levansucrase from Zymomonas mobilis. Ki-Hyo Jang; Ki-Bang Song; Chul Ho Kim; Bong Hyun Chung; Soon Ah Kang; Uck-Han Chun; Ryo Won Choue; Sang-Ki Rhee // Biotechnology Letters;Mar2001, Vol. 23 Issue 5, p339 

  The characteristics of levan formation by different preparations of levansucrase (free and immobilized enzyme and toluene-permeabilized whole cells), derived from recombinant levansucrase from Zymomonas mobilis expressed in Escherichia coli, were investigated. The maximal yield of levan by the...

• Construction of a novel cell-surface display system for heterologous gene expression in Escherichia coli by using an outer membrane protein of Zymomonas mobilis as anchor motif. Ming-xiong He; Hong Feng; Yi-zheng Zhang // Biotechnology Letters;Dec2008, Vol. 30 Issue 12, p2111 

  A novel bacterial cell-surface display system was developed in Escherichia coli using omp1, a hypothetical outer membrane protein of Zymomonas mobilis. By using this system, we successfully expressed β-amylase gene of sweet potato in E. coli. The display of enzyme on the membrane surface was...

• Growth rate of a non-fermentative Escherichia coli strain is influenced by NAD+ regeneration. Vázquez-Limón, Consuelo; Vega-Badillo, Joel; Martínez, Alfredo; Espinosa-Molina, Gabriela; Gosset, Guillermo; Soberón, Xavier; López-Munguía, Agustín; Osuna, Joel // Biotechnology Letters;Dec2007, Vol. 29 Issue 12, p1857 

  By complementing a non-fermentative Escherichia coli ( ldhA − pflB −) strain with the recombinant Zymomonas mobilis ethanol pathway ( pdc, adhB), we evaluated the effect of different levels of enzymatic activity on growth rate demonstrating that there is a direct relationship between...

• Characterization of multiple promoters and transcript stability in the sacB—sacC gene cluster in Zymomonas mobilis. Senthilkumar, V.; Rajendhran, J.; Busby, S. J. W.; Gunasekaran, P. // Archives of Microbiology;Jun2009, Vol. 191 Issue 6, p529 

  In Zymomonas mobilis, the extracellular levansucrase (SacB) and extracellular sucrase (SacC) are involved in sucrose hydrolysis. Genes coding for these two enzymes (s acB and sacC) are arranged in a cluster in the genome and separated by a short intervening sequence. The level of sacC transcript...

Read the Article