Characterization of cold-tolerant trehalose-6-phosphate synthase from the deep-sea bacterium Microbacterium sediminis YLB-01

Figures & data

Figure 1. Expression and purification of recombinant trehalose-6-phosphate synthase (TPS).

Figure 2. Enzymatic properties of recombinant trehalose-6-phosphate synthase (TPS).

Figure 3. Kinetics analysis of recombinant trehalose-6-phosphate synthase (TPS) reaction rates.

Figure 4. Structure analysis of TPS, 1GZ5 and 5K5C.

Figure 5. Maximum-Likelihood tree showing the phylogenetic positions of strain YLB-01^T and other OTSA protein sequences.

Figure 6. Multiple amino acid sequence alignment of TPS and other OTSA.

Table 1. Comparison of the enzymatic properties of trehalose-6-phosphate synthases expressed by various bacteria.

Srtains	Optimum temp (℃)	Heat stability (℃)	Optimum pH	Mental ions	Reference
	TPS	TPS	TPS	TPS
M. sediminis YLB-01	40	72.6	7.5	Mg^2+, Co^2+,Ba^2+	This study
E. coli	40	50	-	-	[14]
M. smegmatis	-	40	7.0	Mg^2+	[6,8]
P. freudenreichii	45	-	5.5	Mg^2+	[12]
R. xylanophilus	60	-	7.0	Mg^2+	[13]
T. thermophilus RQ1	98	100	~6	Not required	[10]
E. halochloris	50	-	7.5	Mg^2+	[7]

Table 2. Comparison of the kinetics of trehalose-6-phosphate synthases expressed by various bacteria.

	TPS
	Km(mM)
Stains	UDPG	G6P	Kcat (S^−1)	Reference
M. sediminis YLB-01	1.38	1.53	58.81/58.46^a	This study
E. coli	8.6	3.7	7.3/6.7^b	[14]
M. smegmatis	0.04	1	-	[6,8]
M. tuberculosis	18	7	-	[12]
P. freudenreichii	0.3	3.0	-	[13]
R. xylanophilus	0.7	2.9	-	[10]
T. thermophilus RQ1	0.5	0.8	176.9	[10]
E. halochloris	1.5	2	-	[7]

a. K_cat of 58.81 S⁻¹ with UDPG as substrate; K_cat of 58.46 S⁻¹ with G6P as substrate.

b. K_cat of 7.3 S⁻¹ with UDPG as substrate; K_cat of 6.7 S⁻¹ with G6P as substrate.

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