A Recombinant Alginate Lyase Algt1 with Potential in Preparing Alginate Oligosaccharides at High-Concentration Substrate
Abstract
:1. Introduction
2. Materials and Methods
2.1. Strains and Chemicals
2.2. Sequence Analysis
2.3. Construction of Recombinant Alginate Lyases
2.4. Heterologous Expression of Alginate Lyases in P. pastoris and Purification
2.5. Enzymatic Activity Assay of Algt and Algt1
2.6. Biochemical Characterization of Algt1
2.7. Analysis on Hydrolysis Products of Algt1
2.8. AOS Batch Preparation at High-Concentration Substrate
3. Results and Discussion
3.1. Sequence Analysis of Algt and Algt1
3.2. Heterologous Expression of Algt and Algt1 in P. pastoris and Their Enzymatic Activity
3.3. Biochemical Properties of Algt1
3.4. Analysis of the Enzymatic Hydrolysates
3.5. Potential of Algt1 for AOS Batch Preparation at High-Concentration Substrate
4. Conclusions
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Alginate Lyase | Source | Family | Expression System | Specific Activity (U/mg) | Optimal Temperature/pH | Thermostability (Remained Enzymatic Activity) | Substrate Concentration | Hydrolytic Products | Ref. |
---|---|---|---|---|---|---|---|---|---|
Algt1 | M. thermotolerans DSM 19189 | PL7 | P. pastoris | 212.86 | 40 °C/7.0 | 80.9% (40 °C, 90 min) | 20% | DP1–4 | This study |
AlgL17 | Microbulbifer sp. ALW1 | PL17 | E. coli | 28.99 | 35 °C/8.0 | 20% (35 °C, 10 min) | 0.5% | DP1–4 | [21] |
Alg2951 | A. portus HB161718T | PL7 | E. coli | / | 25 °C/8.0 | 66.5% (40 °C, 30 min) | 1% | DP1, DP3 | [22] |
AlgSH17 | Microbulbifer sp. SH-1 | PL17 | E. coli | 55.62 | 30 °C/7.0 | 20% (40 °C, 60 min) | 1% | DP1–6 | [38] |
Alg17B | Strain BP-2 | PL17 | Native | 145 | 40–45 °C/7.5–8.0 | 10% (45 °C, 60 min) | 2% | DP1–6 | [37] |
PsMan8A | P. salina | PL8 | P. pastoris | 245 | 25 °C/5.0 | ND | 0.15% | DP1–2 | [23] |
Alys1 | Tamlana sp. s12 | PL7 | E. coli | 1350 | 35 °C/7.0–8.0 | 70% (50 °C, 60 min) | 1% | DP1 | [24] |
VsAly7D | Vibrio sp. QY108 | PL7 | E. coli | 663 | 35 °C/8.0 | 20% (40 °C, 60 min) | 0.3% | DP1–2 | [25] |
Parameters | Alginate | Poly M | Poly G |
---|---|---|---|
Specific activity (U/mg) | 212.86 | 93.87 | 186.19 |
Km (mM) | 0.71 | 1.38 | 0.85 |
Vmax (nmol/s) | 8.56 | 5.83 | 8.09 |
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Liang, Q.; Huang, Y.; Liu, Z.; Xiao, M.; Ren, X.; Liu, T.; Li, H.; Yu, D.; Wang, Y.; Zhu, C. A Recombinant Alginate Lyase Algt1 with Potential in Preparing Alginate Oligosaccharides at High-Concentration Substrate. Foods 2023, 12, 4039. https://doi.org/10.3390/foods12214039
Liang Q, Huang Y, Liu Z, Xiao M, Ren X, Liu T, Li H, Yu D, Wang Y, Zhu C. A Recombinant Alginate Lyase Algt1 with Potential in Preparing Alginate Oligosaccharides at High-Concentration Substrate. Foods. 2023; 12(21):4039. https://doi.org/10.3390/foods12214039
Chicago/Turabian StyleLiang, Qingping, Youtao Huang, Zhemin Liu, Mengshi Xiao, Xinmiao Ren, Tianhong Liu, Hongyan Li, Dongxing Yu, Ying Wang, and Changliang Zhu. 2023. "A Recombinant Alginate Lyase Algt1 with Potential in Preparing Alginate Oligosaccharides at High-Concentration Substrate" Foods 12, no. 21: 4039. https://doi.org/10.3390/foods12214039