Biosynthesis, purification, and biochemical properties of an alkaline-thermostable keratinase derived from Bacillus cereus J6

Keratinase production by B. cereus J6. A. Enzyme production profile; B, Feather; C, Nitrogen sources; D, Urea; E, Carbon sources; F, Lactose; G, Mineral salts; F, MgSO4.

Impact of cultural conditions on B. cereus J6 producing keratinase

SDS-PAGE analysis of the purification of J6 keratinase. Lane M, protein marker; Lane 1, the crude B. cereus J6 keratinase; Lane 2, ASP; Lane 3, the target enzyme.

The enzymatic activity of the keratinase towards various substrates

Effects of chemicals on J6 keratinase

Chemicals	Concentration	Relative activity (%)
Na+	5 mmol/L	102.59±2.78
K⁺	5 mmol/L	99.03±2.93
Mn²⁺	5 mmol/L	106.73±2.25
Ca²⁺	5 mmol/L	122.56±2.19
Mg²⁺	5 mmol/L	131.81±5.14
Zn²⁺	5 mmol/L	118.52±3.52
Cu²⁺	5 mmol/L	60.87±4.53
Al³⁺	5 mmol/L	62.67±6.21
Fe²⁺	5 mmol/L	52.89±5.35
Fe³⁺	5 mmol/L	31.04±3.87
Tween 20	1%	115.67±3.14
Tween 60	1%	175.27±1.31
Tween 80	1%	150.16±2.31
Triton X-100	1%	106.06±0.52
SDS	1%	84.30±2.56
DTT	1 mmol/L	155.96±2.73
EDTA	1 mmol/L	5.05±2.21
PMSF	1 mmol/L	Not detected

Results of orthogonal experiment

Number	A	B	C	D	Keratinase yield (U/mL)
1	1	1	1	1	375.55±3.12
2	1	2	2	2	179.8±5.93
3	1	3	3	3	359.9±6.49
4	2	1	2	3	401.4±9.84
5	2	2	3	1	342.55±3.16
6	2	3	1	2	208.9±9.53
7	3	1	3	2	186.9±42
8	3	2	1	3	307.25±4.78
9	3	3	2	1	294.9±2.65
K1	305.08	321.28	297.23	337.67
K2	317.62	276.53	292.03	192.86
K3	263.02	288.9	296.45	356.18
R	54.6	32.38	5.2	163.32
Order of priority: D > A > B > C
Optimal Combination: A₂B₁C₁D₃

Orthogonal experimental design of four factors and three levels

Level	Factor
Level	A feather (g/L)	B Lactose (g/L)	C Urea (g/L)	D MgSO4 (g/L)
1	7.5	2.5	7.5	0.9
2	10	5	10	1
3	12.5	7.5	12.5	1.1

Purification of the keratinase produced by B_ cereus J6

Procedures	Protein quantity (mg)	activity (U)	Specific activity (U/mg)	Purification fold	Recovery rate (%)
Supernatant	53.25	24032	451.31	1.00	100
ASP	15.41	14256	925.19	2.05	59.32
DEAE-650 M chromatography	0.65	2739	4215.24	9.34	11.36

Sprache:: Englisch

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Fachgebiete der Zeitschrift:: Biologie, Biologie, andere, Mathematik, Angewandte Mathematik, Mathematik, Allgemeines, Physik, Physik, andere

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Biosynthesis, purification, and biochemical properties of an alkaline-thermostable keratinase derived from Bacillus cereus J6

Online veröffentlicht: 17. März 2025

Eingereicht: 30. Okt. 2024

Akzeptiert: 10. Feb. 2025

DOI: https://doi.org/10.2478/amns-2025-0831

Schlüsselwörter
<kwd>keratinase</kwd>, <kwd><italic>Bacillus cereus</italic></kwd>, <kwd>production</kwd>, <kwd>purification</kwd>

© 2025 Rongxian Zhang et al., published by Sciendo

This work is licensed under the Creative Commons Attribution 4.0 International License.

Figure 1.