Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (8): 283-290.doi: 10.13560/j.cnki.biotech.bull.1985
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RAO Zi-huan(), XIE Zhi-xiong()
Received:
2023-01-09
Online:
2023-08-26
Published:
2023-09-05
Contact:
XIE Zhi-xiong
E-mail:RZHmio@whu.edu.cn;zxxie@whu.edu.cn
RAO Zi-huan, XIE Zhi-xiong. Isolation and Identification of a Cellulose-degrading Strain of Olivibacter jilunii and Analysis of Its Degradability[J]. Biotechnology Bulletin, 2023, 39(8): 283-290.
Fig. 4 Genome-wide covariance alignment of strain 18B and O. jilunii 14-2AT It in the upper part of the figure it is strain 18B and in the lower part is O. jilunii 14-2AT
Fig. 5 Genome alignment within the genus Olivibacter Complete genome sequence information of related strains in Olivibacter genus was downloaded from NCBI and compared with strain 18B. In the figure, sitiens indicates the strain O. sitiensis AW-6T, domesticus indicates O. domesticus DC186T, and LS_1 indicates O. sp.LS-1. jilunii refers to strain O. jilunii 14-2AT
项目 Item | 18B | O. jilunii 14-2AT |
---|---|---|
生长温度/℃ | 12-48 | 4-42 |
生长pH | 6.0-8.0 | 6.0-9.0 |
NaCl耐受/% | 0-6 | 0-5 |
氧化酶 | + | - |
硝酸盐还原 | + | + |
糖酵解 | - | + |
VP | + | + |
β-半乳糖苷酶 | + | + |
革兰氏染色 | - | - |
MacConkey agar | +(Red) | +(Red) |
Table 1 Comparison of common physiological and bioche-mical characteristics between 18B and O. jilunii 14-2AT
项目 Item | 18B | O. jilunii 14-2AT |
---|---|---|
生长温度/℃ | 12-48 | 4-42 |
生长pH | 6.0-8.0 | 6.0-9.0 |
NaCl耐受/% | 0-6 | 0-5 |
氧化酶 | + | - |
硝酸盐还原 | + | + |
糖酵解 | - | + |
VP | + | + |
β-半乳糖苷酶 | + | + |
革兰氏染色 | - | - |
MacConkey agar | +(Red) | +(Red) |
Fig. 6 Variation of 18B cellulase activity with incubation time The gray line is the cellulase activity of the strain after transfer from CMC-Na culture environment, and the red line is the cellulase activity of the strain after transfer from CMC-Na culture environment to LB culture for one generation. The enzyme activity of O. jilunii 14-2AT was not detected
Fig. 7 Gene amplification after induction of expression Marker is Trans2k plusII. D is the precursor gene of endoglucanase D, the length is 1 812 bp. M is cellulase M gene with length of 1 119 bp. Z is the precursor gene of endoglucanase Z, the length of which is 1 542 bp. P is a pure endoglucanase gene with a length of 1 395 bp
Fig. 8 Protein assay of cellulase gene induced expression 0: Maker. 1: The whole solution broken after D protein expression; 2: the supernatant broken after D protein expression; 3: the precipitation broken after D protein expression; 4: the whole fluid broken after Z protein expression; 5: the supernatant broken after Z protein expression; 6: the precipitation broken after Z protein expression; 7: the whole solution of cell breaking after no-load induction expression; 8: the supernatant of cell breaking after no-load induction expression; 9: the precipitation of cell breaking after no-load induction expression; 10: marker; 11: the whole solution broken after M protein expression; 12: the supernatant broken after M protein expression; 13: the precipitation broken after M protein expression; 14: the whole fluid broken after P protein expression; 15: the supernatant broken after P protein expression; 16: the precipitation broken after P protein expression; 17: the whole solution of cell breaking after no-load induction expression; 18: the supernatant of cell breaking after no-load induction expression; 19: the precipitation of cell breaking after no-load induction expression. The red box marked the target protein region
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