PMID- 18712537
OWN - NLM
STAT- MEDLINE
DCOM- 20090306
LR  - 20211020
IS  - 1367-5435 (Print)
IS  - 1367-5435 (Linking)
VI  - 35
IP  - 11
DP  - 2008 Nov
TI  - Expression, purification and immobilization of the intracellular invertase INVA, 
      from Zymomonas mobilis on crystalline cellulose and Nylon-6.
PG  - 1455-63
LID - 10.1007/s10295-008-0447-1 [doi]
AB  - This paper presents two immobilization methods for the intracellular invertase 
      (INVA), from Zymomonas mobilis. In the first method, a chimeric protein 
      containing the invertase INVA, fused through its C-terminus to CBDCex from 
      Cellulomonas fimi was expressed in Escherichia coli strain BL21 (DE3). INVA was 
      purified and immobilized on crystalline cellulose (Avicel) by means of affinity, 
      in a single step. No changes were detected in optimal pH and temperature when 
      INVA-CBD was immobilized on Avicel, where values of 5.5 and 30 degrees C, 
      respectively, were registered. The kinetic parameters of the INVA-CBD fusion 
      protein were determined in both its free form and when immobilized on Avicel. Km 
      and Vmax were affected with immobilization, since both showed an increase of up 
      to threefold. Additionally, we found that subsequent to immobilization, the 
      INVA-CBD fusion protein was 39% more susceptible to substrate inhibition than 
      INVA-CBD in its free form. The second method of immobilization was achieved by 
      the expression of a 6xHis-tagged invertase purified on Ni-NTA resin, which was 
      then immobilized on Nylon-6 by covalent binding. An optimal pH of 5.5 and a 
      temperature of 30 degrees C were maintained, subsequent to immobilization on 
      Nylon-6 as well as with immobilization on crystalline cellulose. The kinetic 
      parameters relating to Vmax increased up to 5.7-fold, following immobilization, 
      whereas Km increased up to 1.7-fold. The two methods were compared showing that 
      when invertase was immobilized on Nylon-6, its activity was 1.9 times that when 
      immobilized on cellulose for substrate concentrations ranging from 30 to 390 mM 
      of sucrose.
FAU - de Los Angeles Calixto-Romo, Maria
AU  - de Los Angeles Calixto-Romo M
AD  - Departamento de Biotecnologia y Bioingenieria, CINVESTAV-IPN. Av. Instituto 
      Politecnico, Nacional 2508 Col. San Pedro Zacatenco, CP 07360, Mexico D.F., 
      Mexico.
FAU - Santiago-Hernandez, Jose Alejandro
AU  - Santiago-Hernandez JA
FAU - Vallejo-Becerra, Vanessa
AU  - Vallejo-Becerra V
FAU - Amaya-Delgado, Lorena
AU  - Amaya-Delgado L
FAU - del Carmen Montes-Horcasitas, Maria
AU  - del Carmen Montes-Horcasitas M
FAU - Hidalgo-Lara, Maria Eugenia
AU  - Hidalgo-Lara ME
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20080820
PL  - Germany
TA  - J Ind Microbiol Biotechnol
JT  - Journal of industrial microbiology & biotechnology
JID - 9705544
RN  - 0 (Enzymes, Immobilized)
RN  - 0 (Polymers)
RN  - 25038-54-4 (nylon 6)
RN  - 6879X594Z8 (Caprolactam)
RN  - 9004-34-6 (Cellulose)
RN  - EC 3.2.1.26 (beta-Fructofuranosidase)
SB  - IM
MH  - Caprolactam/analogs & derivatives/chemistry
MH  - Cellulose/chemistry
MH  - Enzyme Stability
MH  - Enzymes, Immobilized/*chemistry/genetics/*isolation & purification/metabolism
MH  - Escherichia coli/genetics/metabolism
MH  - *Gene Expression
MH  - Kinetics
MH  - Polymers/chemistry
MH  - Zymomonas/*enzymology
MH  - beta-Fructofuranosidase/*chemistry/genetics/*isolation & purification/metabolism
EDAT- 2008/08/21 09:00
MHDA- 2009/03/07 09:00
CRDT- 2008/08/21 09:00
PHST- 2008/04/23 00:00 [received]
PHST- 2008/07/30 00:00 [accepted]
PHST- 2008/08/21 09:00 [pubmed]
PHST- 2009/03/07 09:00 [medline]
PHST- 2008/08/21 09:00 [entrez]
AID - 10.1007/s10295-008-0447-1 [doi]
PST - ppublish
SO  - J Ind Microbiol Biotechnol. 2008 Nov;35(11):1455-63. doi: 
      10.1007/s10295-008-0447-1. Epub 2008 Aug 20.