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Study on oligomerization of glutamate decarboxylase from Lactobacillus brevis using asymmetrical flow field-flow fractionation (AF4) with light scattering techniques

Choi, Jaeyeong; Lee, Seungho LU ; Linares-Pastén, Javier A. LU and Nilsson, Lars LU (2018) In Analytical and Bioanalytical Chemistry 410(2). p.451-458
Abstract

In this work, asymmetrical flow field-flow fractionation (AF4) coupled with UV/Vis, multi-angle light scattering (MALS), and differential refractive index (dRI) detectors (AF4-UV-MALS-dRI) was employed for analysis of glutamate decarboxylase (LbGadB) from Lactobacillus brevis (L. brevis). AF4 provided molecular weight (MW) (or size)-based separation of dimer, hexamer, and aggregates of LbGadB. The effect of pH on oligomerization of LbGadB was investigated, and then AF4 results were compared to those from molecular modeling. The MWs measured by AF4-UV-MALS-dRI for dimeric and hexameric forms of LbGadB were 110 and 350 kDa, respectively, which are in good agreements with those theoretically calculated (110 and 330 kDa). The molecular... (More)

In this work, asymmetrical flow field-flow fractionation (AF4) coupled with UV/Vis, multi-angle light scattering (MALS), and differential refractive index (dRI) detectors (AF4-UV-MALS-dRI) was employed for analysis of glutamate decarboxylase (LbGadB) from Lactobacillus brevis (L. brevis). AF4 provided molecular weight (MW) (or size)-based separation of dimer, hexamer, and aggregates of LbGadB. The effect of pH on oligomerization of LbGadB was investigated, and then AF4 results were compared to those from molecular modeling. The MWs measured by AF4-UV-MALS-dRI for dimeric and hexameric forms of LbGadB were 110 and 350 kDa, respectively, which are in good agreements with those theoretically calculated (110 and 330 kDa). The molecular sizes determined by AF4-UV-MALS-dRI were also in good agreement with those obtained from molecular modeling (6 and 10 nm, respectively, for dimeric and hexameric from AF4-UV-MALS-dRI and 6.4 × 7.6 and 7.6 × 13.1 nm from molecular modeling). The effects of temperature, salt type, and salt concentration on oligomerization of LbGadB were also investigated using dynamic light scattering (DLS). It was found that the hexameric form of LbGadB was most stable at pH 6 and in presence of NaCl or KCl. The results indicate that AF4, in combination of various online detectors mentioned above, provides an effective tool for monitoring of oligomerization of LbGadB under different conditions, such as temperature, pH, type of salts, and salt concentrations.

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organization
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Contribution to journal
publication status
published
subject
keywords
Asymmetrical flow field-flow fractionation (AF4), Glutamate decarboxylase (GAD), Lactobacillus brevis (L. brevis), Multi-angle light scattering (MALS), Oligomerization, Probiotic
in
Analytical and Bioanalytical Chemistry
volume
410
issue
2
pages
451 - 458
publisher
Springer
external identifiers
  • scopus:85034622007
ISSN
1618-2642
DOI
10.1007/s00216-017-0735-6
language
English
LU publication?
yes
id
1072cb71-6f34-4792-bfbc-b460f35f0e05
date added to LUP
2017-12-08 07:25:18
date last changed
2018-03-10 03:00:07
@article{1072cb71-6f34-4792-bfbc-b460f35f0e05,
  abstract     = {<p>In this work, asymmetrical flow field-flow fractionation (AF4) coupled with UV/Vis, multi-angle light scattering (MALS), and differential refractive index (dRI) detectors (AF4-UV-MALS-dRI) was employed for analysis of glutamate decarboxylase (LbGadB) from Lactobacillus brevis (L. brevis). AF4 provided molecular weight (MW) (or size)-based separation of dimer, hexamer, and aggregates of LbGadB. The effect of pH on oligomerization of LbGadB was investigated, and then AF4 results were compared to those from molecular modeling. The MWs measured by AF4-UV-MALS-dRI for dimeric and hexameric forms of LbGadB were 110 and 350 kDa, respectively, which are in good agreements with those theoretically calculated (110 and 330 kDa). The molecular sizes determined by AF4-UV-MALS-dRI were also in good agreement with those obtained from molecular modeling (6 and 10 nm, respectively, for dimeric and hexameric from AF4-UV-MALS-dRI and 6.4 × 7.6 and 7.6 × 13.1 nm from molecular modeling). The effects of temperature, salt type, and salt concentration on oligomerization of LbGadB were also investigated using dynamic light scattering (DLS). It was found that the hexameric form of LbGadB was most stable at pH 6 and in presence of NaCl or KCl. The results indicate that AF4, in combination of various online detectors mentioned above, provides an effective tool for monitoring of oligomerization of LbGadB under different conditions, such as temperature, pH, type of salts, and salt concentrations.</p>},
  author       = {Choi, Jaeyeong and Lee, Seungho and Linares-Pastén, Javier A. and Nilsson, Lars},
  issn         = {1618-2642},
  keyword      = {Asymmetrical flow field-flow fractionation (AF4),Glutamate decarboxylase (GAD),Lactobacillus brevis (L. brevis),Multi-angle light scattering (MALS),Oligomerization,Probiotic},
  language     = {eng},
  number       = {2},
  pages        = {451--458},
  publisher    = {Springer},
  series       = {Analytical and Bioanalytical Chemistry},
  title        = {Study on oligomerization of glutamate decarboxylase from Lactobacillus brevis using asymmetrical flow field-flow fractionation (AF4) with light scattering techniques},
  url          = {http://dx.doi.org/10.1007/s00216-017-0735-6},
  volume       = {410},
  year         = {2018},
}