Isolation, Characterization, and Metabolic Profiling of Ceratorhiza hydrophila from the Aquatic Plant Myriophyllum spicatum

Figures & data

Figure 1. Morphological and microscopic examination of Ceratorhiza hydrophila. (a) growth on Potato dextrose agar media after seven days of incubation at 25ºC, (b) brown globose sclerotia (4X), (c) septate branched mycelium (40×).

Figure 2. Maximum likelihood tree showed that Ceratorhiza hydrophila isolate EG19 (MK387081) had a high similarity of 99% toward Ceratorhiza hydrophila isolate SR6 (JQ359765).

Figure 3. Ceratorhiza hydrophila enzymatic activities after incubation for 72 h at 25ºC. (a) protease, (b) cellulase.

Table 1. Enzyme production by Ceratorhiza hydrophila.

Enzyme	Clear zone diameter (cm)
Protease	3.6 ± 0.25^b
Asparaginase	0.0^c
Cellulase	5.5 ± 0.1^a
Amylase	0.0^c

In each column means followed by the same letter are un-significantly at p < 0.05 based on Fisher test.

Table 2. Antibacterial activity of the crude extracts of Ceratorhiza hydrophila against different tested bacterial strains.

Bacterial strain	Inhibition zone (mm)
	C. hydrophila (55 mg ml^−1 )	Clindamycin (2 m µg) (Positive control)	Ciprofloxacin (5 m µg) (Positive control)	Ethyl acetate (Negative control)
Streptococcus pneumonia	18 ± 1.4^ab	15 ± 0.06	ND	0 ± 0
Micrococcus luteus	20 ± 0.7^a	24 ± 0.06	20 ± 0.0	0 ± 0
Staphylococcus aureus	16 ± 0.7^b	22 ± 0.06	30 ± 0.0	0 ± 0
Escherichia coli	17 ± 2.1^b	ND	25±.1	0 ± 0
Proteus sp.	0 ± 0^c	ND	22 ± 0.1	0 ± 0

In each column means followed by the same letter are un-significantly at p < 0.05 based on Fisher test. Antibacterial activity is presented as means of measurements of inhibition zones (mm) ± SD. ND means not detected.

Figure 4. ¹H NMR spectra of the polar extract of Ceratorhiza hydrophila. (a) Aliphatic region, (b) sugars abundance signals region, (c) aromatic region.

Table 3. The identified compounds in the polar extract of Ceratorhiza hydrophila with their chemical formulas, chemical shifts, multiplicity and coupling constants.

	Compound name	Chemical formula	^1H (ppm) (functional group or specific H, multiplicity)	^13C (ppm) (functional group or specific C)	J (Hz)
1	Acetate	C2H4O2	1.92 (CH3, s)	25.6 (CH3)	–
2	Alanine	C3H7NO2	3.77 (H^α, q)	53.5 (C^α)	7.20
			1.48 (H^β, d)	18.6 (C^β)	7.25
3	Betaine	C5H11NO2	3.28 (CH3, s)	56.2 (CH3)	–
			3.90 (CH2, s)	69.2 (CH2)	–
4	Choline	C5H14NO	4.05 (H^α ,m)	58.6 (C^α)	–
			3.21 (H^γ, s)	56.8 (C^γ)	–
			3.50 (H^β, m)	70.4 (C^β)	–
5	Dimethylamine	C2H7N	2.72 (CH3, s)	37.0 (CH3)	–
6	Formate	CH2O2	8.46 (CH, s)	–	–
7	Fumarate	C4H4O4	6.52 (CH, s)	137.3 (CH)	–
8	Glucose	C6H12O6	4.63 (^1βCH, d)	98.9 (^1αCH)	7.88
			5.24 (^1αCH, d)	94.7 (^1βCH)	3.67
			3.47 (^2αCH, m)	74.4 (^2αCH)
			3.39 (^4CH, m)	72.6 (^4CH)
			3.70 (^5βCH, m)	63.7 (^6CH)
9	Glycerol	C3H8O3	3.57 (CH2, dd)	65.3 (CH2)	6.25,6.65
			3.65 (CH2, dd)	65.4 (CH2)
10	Glycine	C2H5NO2	3.55 (H^α, s)	44.3 (C^α)	–
11	Isoleucine	C6H13NO2	1.26 (H^γ, m)	26.8 (C^γ)	–
			1.01 (H^γ, d)	17.5 (C^γ)	7.0
			0.92 (H^δ, t)	–	7.1
12	Lactate	C3H6O3	1.33 (CH3, d)	22.1 (CH3)	6.87
13	Leucine	C6H13NO2	0.97 (H^δ, t)	24.0 (C^δ)	6.09
			1.07 (H^β, m)	–	–
14	Malate	C4H6O5	2.34 (CH2, dd)	45.0 (CH2)	15.37,10.2
			2.68 (CH2, dd)	45.2 (CH2)	15.37,2.9
15	Malonate	C3H4O4	3.12 (H^β, s)	50.2 (C^β)	–
16	O-Phosphocholine	C5H15NO4P	3.20 (CH3, s)	56.7 (CH3)	–
			3.57 (CH2, m)	68.9 (CH2)	–
			4.15 (CH2, m)	60.6 (CH2)	–
17	Pyruvate	C3H4O3	2.38 (CH3, s)	29.3 (CH3)	–
18	Succinate	C4H6O4	2.41 (CH2, s)	36.4 (CH2)	–
19	Threonine	C4H9NO3	1.31 (CH3, d)	22.5 (C^α)	6.49
			3.58 (CH3, d)	63.4 (C^γ)	5.20
20	Trehalose	C12H22O11	5.20 (^1CH, d)	95.8 (^1CH)	3.73
			3.66 (^2CH, dd)	73.6 (^2CH)	10,3.97
			3.46 (CH, t)		9.54
21	Trimethylamine	C3H9N	2.90 (CH3, s)	47.4 (CH3)	–
22	Valine	C5H11NO2	3.60 (H^α, d)	63.0 (C^α)	4.54
			0.99 (H^γ, d)	20.8 (C^γ)	7.00
			1.04 (H^γ, d)	20.4 (C^γ)	7.00
23	Xanthine	C5H4N4O2	7.96 (CH, s)	140.3 (CH)	–