Immobilization of β-Galactosidases from <i>Lactobacillus</i> on Chitin Using a Chitin-Binding Domain

Pham, Mai-Lan; Leister, Tatjana; Nguyen, Hoang Anh; Do, Bien-Cuong; Pham, Anh-Tuan; Haltrich, Dietmar; Yamabhai, Montarop; Nguyen, Thu‐Ha; Nguyen, Tien‐Thanh

doi:10.1021/acs.jafc.6b04982

Cited by 27 publications

(19 citation statements)

References 42 publications

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“…However, the V max (14.57 nmol/min/µg) and K cat (0.07 min −1 ) of I-ChBD-CadA were lower than that of ChBD-CadA, which indicated that the activity of ChBD-CadA decreased after immobilization. Pham et al (2017) also found the activity of β-galactosidases declined after immobilization on chitin, which was similar with our study.…”

Section: The Effect Of Chbd On 3d Structure Of Cadasupporting

confidence: 92%

“…These results suggested that the fusion of ChBD affected the expression of CadA gene. Other studies also reported that the gene expression will decline after fusion of ChBD or other genes (Pham et al, 2017), which are similar with this study.…”

Section: Construction and Expression Of Fusion Genesupporting

confidence: 91%

“…These results suggest that the majority of ChBD-CadA can be efficiently bound to chitin due to the high specific affinity of ChBD for chitin. Other studies also reported that ChBD possesses significant binding affinity for various enzyme immobilizations (Pham et al, 2017;Xu et al, 2017).…”

Section: Simultaneous Purification and Immobilization Of Chbd-cada Bymentioning

confidence: 89%

“…Based on the above results, the optimal immobilized conditions were as follows: temperature, 25 • C; time, 10 min; pH, 6.2; protein concentration, 1.20 mg/mL, which can bind with CadA activity of 68.67 U/mL and 93% immobilized ratio. Pham et al (2017) also reported that the immobilized ratio of fusion protein can reach 99% and 91.0% using the ChBD of chitinase A1, respectively.…”

Section: Simultaneous Purification and Immobilization Of Chbd-cada Bymentioning

confidence: 94%

“…The chitin-binding domain (ChBD) is a portion of some chitin hydrolysis enzymes and can specifically bind to chitin (the second most abundant natural polysaccharide after cellulose) via an affinity tag, which makes ChBD a popular choice for enzyme immobilization via protein fusion (Hashimoto et al, 2000;Simsek et al, 2013). ChBD has been employed to immobilize trehalose, levansucrase, β-galactosidase, and heparinase on chitin with excellent enzymatic outcomes (Chiang et al, 2009;Pham et al, 2017;Xu et al, 2017;Jiang et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

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Cadaverine Production From L-Lysine With Chitin-Binding Protein-Mediated Lysine Decarboxylase Immobilization

Zhou

Zhang

Wei

et al. 2020

Front. Bioeng. Biotechnol.

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Lysine decarboxylase (CadA) can directly convert L-lysine to cadaverine, which is an important platform chemical that can be used to produce polyamides. However, the non-recyclable and the poor pH tolerance of pure CadA hampered its practical application. Herein, a one-step purification and immobilization procedure of CadA was established to investigate the cadaverine production from L-lysine. Renewable biomass chitin was used as a carrier for lysine decarboxylase (CadA) immobilization via fusion of a chitin-binding domain (ChBD). Scanning electron microscopy, laser scanning confocal microscopy, fourier transform infrared spectra, elemental analysis, and thermal gravimetric analysis proved that the fusion protein ChBD-CadA can be adsorbed on chitin effectively. Furthermore, the fusion protein (ChBD-CadA) existed better pH stability compared to wild CadA, and kept over 73% of the highest activity at pH 8.0. Meanwhile, the ChBD-CadA showed high specificity toward chitin and reached 93% immobilization yield within 10 min under the optimum conditions. The immobilized ChBD-CadA (I-ChBD-CadA) could efficiently converted L-lysine at 200.0 g/L to cadaverine at 135.6 g/L in a batch conversion within 120 min, achieving a 97% molar yield of the substrate L-lysine. In addition, the I-ChBD-CadA was able to be reused under a high concentration of L-lysine and retained over 57% of its original activity after four cycles of use without acid addition to maintain pH. These results demonstrate that immobilization of CadA using chitin-binding domain has the potential in cadaverine production on an industrial scale.

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Section: The Effect Of Chbd On 3d Structure Of Cadasupporting

confidence: 92%

Section: Construction and Expression Of Fusion Genesupporting

confidence: 91%

Section: Simultaneous Purification and Immobilization Of Chbd-cada Bymentioning

confidence: 89%

Section: Simultaneous Purification and Immobilization Of Chbd-cada Bymentioning

confidence: 94%

Section: Introductionmentioning

confidence: 99%

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Cadaverine Production From L-Lysine With Chitin-Binding Protein-Mediated Lysine Decarboxylase Immobilization

Zhou

Zhang

Wei

et al. 2020

Front. Bioeng. Biotechnol.

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Immobilization of Microbial Consortium for Glutaric Acid Production from Lysine

et al. 2021

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Glutaric acid is an important C5 platform compound for the synthesis of nylon polymers. However, its production titer and cost are limited by the need for expensive cofactors addition as well as the difficulty of reusing microbial catalysts. Herein, to address these problems, two regeneration systems each for NAD(P) + and α-ketoglutaric acid were introduced into a fiveenzyme cadaverine pathway for a whole-cell biocatalytic approach. We demonstrated the efficient conversion of lysine to glutaric acid in a synthetic microbial consortium manner, producing 73.2 g/L glutaric acid with a 74 % molar yield in a 5-L bioreactor scale. Moreover, with a colloidal chitin-based immobilization, the engineered microbial consortium exhibited good stability and reusability in a five-cycle bioconversion. This study provides an efficient microbial platform for glutaric acid production by developing a cheaper and universal cell immobilization approach.

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Elucidating the binding efficacy of β‐galactosidase on polyaniline–chitosan nanocomposite and polyaniline–chitosan–silver nanocomposite: activity and molecular docking insights

Khan

Husain

Ahmad

2018

J of Chemical Tech & Biotech

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BACKGROUND: Aspergillus oryzae -galactosidase ( -gal; EC 3.2.1.23) was physically adsorbed on polyaniline-chitosan nanocomposite (PANI-CS-NC) and polyaniline-chitosan-silver nanocomposite (PANI-CS-Ag-NC). Stability and kinetic studies were assessed, along with reusability and molecular docking interactions.RESULTS: According to transmission electron microscopy images, the sizes of the respective NCs were found to be in the range of 23-39 and 6-18 nm. The (effectiveness factor) values for PANI-CS-NC-and PANI-CS-Ag-NC-bound -gal were calculated to be 0.89 and 1.23, respectively, suggesting an enhancement in the activity of the enzyme in the presence of silver ions. Significant broadening was observed in pH and temperature profiles. Kinetic properties of PANI-CS-NC-and PANI-CS-Ag-NC-bound -gal showed lower K m , and two-and fourfold higher V max , respectively, compared to free enzyme. PANI-CS-Ag-NC-bound -gal was able to retain 94% of its initial activity after 10 repeated uses. Circular dichroism analysis revealed conformational changes in the secondary structure of protein upon immobilization. The genotoxic assessment revealed non-toxic behaviour of both free and enzyme-bound NC. The rate of lactose hydrolysis at 50 ∘ C exhibited by PANI-CS-Ag-NC bound -gal was 7% higher than that for the native enzyme. In-silico analysis showed efficient binding of -gal on PANI-CS-NCs due to interaction with residues that were exclusive of the active site of the enzyme. CONCLUSION: PANI-CS-Ag-NC-bound -gal preparation proved to be a robust nano-biocatalyst due to remarkable enhancement in catalytic activity along with excellent stability and reusability characteristics, which could be further utilized for efficient construction of nano-biosensors for producing lactose-free dairy products to feed lactose-intolerant patients. Lactose hydrolysis in batch processEnzymatic hydrolysis of lactose is a highly selective procedure which maintains the characteristics of a raw material in the J Chem Technol Biotechnol 2019; 94: 837-849

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Immobilization of β-Galactosidases from Lactobacillus on Chitin Using a Chitin-Binding Domain

Cited by 27 publications

References 42 publications

Cadaverine Production From L-Lysine With Chitin-Binding Protein-Mediated Lysine Decarboxylase Immobilization

Cadaverine Production From L-Lysine With Chitin-Binding Protein-Mediated Lysine Decarboxylase Immobilization

Immobilization of Microbial Consortium for Glutaric Acid Production from Lysine

Elucidating the binding efficacy of β‐galactosidase on polyaniline–chitosan nanocomposite and polyaniline–chitosan–silver nanocomposite: activity and molecular docking insights

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