Potential pancreatic lipase inhibitory activity of an endophytic Penicillium species

Figures & data

Table 1. Endophytic fungi from different medicinal plants used during the study with their corresponding inhibition activities in two chromogenic in vitro qualitative assays.

Culture code	Tentative identification	Host plant	Part of the plant	Rhodamine assay	Phenol red assay
#1 CCBD	Fusarium sp	Cinnamomum camphora	Bark	−−	−−
#1 CCSTITD	Muscodor sp	C. camphora	Stem internal tissue	++	++
#5 CZBAWLS	Fusarium sp	Cinnamomum zeylanicum	Bark	+	+
#2104 CZSTITG	Alternaria sp	C. zeylanicum	Stem internal tissue	−−	−−
#2106 CZSTITG	Curvularia sp	C. zeylanicum	Stem internal tissue	−−	−−
#2112 CZSTITG	Nigro spora sp	C. zeylanicum	Stem internal tissue	−−	−−
#2117 CZSTITG	Alternaria sp	C. zeylanicum	Stem internal tissue	−−	−−
#2125 CZSTITG	Unidentified	C. zeylanicum	Stem internal tissue	−−	−−
#2131 CZSTITG	Unidentified	C. zeylanicum	Stem internal tissue	−−	−−
#2143 CZSTITG	Alternaria sp	C. zeylanicum	Stem internal tissue	−−	−−
#2158 CZSTITG	Pestalotiopsis sp	C. zeylanicum	Stem internal tissue	−−	−−
#2161 CZSTITG	Unidentified	C. zeylanicum	Stem internal tissue	−−	−−
#2162 CZSTITG	Aspergillus sp	C. zeylanicum	Stem internal tissue	+	++
#2168 CZSTITG	Aspergillus sp	C. zeylanicum	Stem internal tissue	−−	−−
# 1 CSSTOT	Schizophyllum sp	Camellia sinensis	Stem	+++	+++
# 2 CSSTOT	Schizophyllum sp	C. sinensis	Stem	++	+++
# 7 CSSTOT	Colletotrichum sp	C. sinensis	Stem	+	++
#16 NOBASVNP	Fusarium sp	Nerium oleander	Bark	++	+
#1 PNLNEY	Alternaria sp	Piper nigrum	Leaf	−−	−−
#2 PNLNEY	Alternaria sp	P. nigrum	Leaf	+	+
#8 PNLNEY	Alternaria sp	P. nigrum	Leaf	++	+++
#14 RSBANEY	Fusarium sp	Rauwolfia serpentina	Bark	−−	−−
#16 RSLBRT	Fusarium sp	R. serpentina	Leaf	+	+
#2 TMSDTYEL	Fusarium sp	Tabernaemontana divaricata	Stem	−	−
#4 TMSDTYEL	Fusarium sp	T. divaricata	Stem	+	+
#7 TBBALM	Mycelia sterile	Taxus baccata	Bark	++	++
#8 TBBALM	Pestalotiopsis sp	T. baccata	Bark	−−	−−
#11 TBBALM	Unidentified	T. baccata	Bark	+	+
#13 TBBALM	Unidentified	T. baccata	Bark	−−	−−
#14 TBBALM	Unidentified	T. baccata	Bark	+	+
#17 TBBALM	Fusarium sp	T. baccata	Bark	−−	+
#20 TBBALM	Alternaria sp	T. baccata	Bark	++	++
#21 TBBALM	Phomopsis sp	T. baccata	Bark	−−	−−
#22 TBBALM	Unidentified	T. baccata	Bark	−−	−−
#25 TBBALM	Phomopsis sp	T. baccata	Bark	+	+
#26 TBBALM	Unidentified	T. baccata	Bark	−−	−−
#27 TBBALM	Unidentified	T. baccata	Bark	−−	−−
#28 TBBALM	Unidentified	T. baccata	Bark	−−	−−
#30 TBBALM	Unidentified	T. baccata	Bark	−−	−−
#31 TBBALM	Ascobolus sp	T. baccata	Bark	++	++
#33 TBBALM	Mycelia sterilia	T. baccata	Bark	+++	+++
#35 TBBALM	Phomopsis sp	T. baccata	Bark	++	++
#36 TBBALM	Alternaria sp	T. baccata	Bark	+	+
#40 TBBALM	Pestalotiopsis sp	T. baccata	Bark	−−	++
#41 TBBALM	Pestalotiopsis sp	T. baccata	Bark	−−	−−
#42 TBBALM	Pestalotiopsis sp	T. baccata	Bark	+	+
#43 TBBALM	Fusarium sp	T. baccata	Bark	−−	−−
#44 TBBALM	Unidentified	T. baccata	Bark	−−	−−
#46 TBBALM	Ascobolus sp	T. baccata	Bark	+	+
#48 TBBALM	Unidentified	T. baccata	Bark	−−	−−
#50 TBBALM	Unidentified	T. baccata	Bark	++	++
#53 TBBALM	Unidentified	T. baccata	Bark	−−	−−
#54 TBBALM	Nectria sp	T. baccata	Bark	++	++
#57 TBBALM	Penicillium sp	T. baccata	Bark	+++	++++
#65 TBBALM	Pestalotiopsis sp	T. baccata	Bark	−−	−−
#67 TBBALM	Unidentified	T. baccata	Bark	+	+
#69 TBBALM	Unidentified	T. baccata	Bark	−−	−−
#73 TBBALM	Unidentified	T. baccata	Bark	−−	−−
#80 TBBALM	Unidentified	T. baccata	Bark	−−	−−
#89 TBBALM	Unidentified	T. baccata	Bark	++	+
#91 TBBALM	Phomopsis sp	T. baccata	Bark	++	+
#93 TBBALM	Pestalotiopsis sp	T. baccata	Bark	++	++
#94 TBBALM	Unidentified	T. baccata	Bark	−−	−−
#98 TBBALM	Penicillium sp	T. baccata	Bark	+	+
#100 TBBALM	Penicillium sp	T. baccata	Bark	−−	−−
#113 TBBALM	Pestalotiopsis sp	T. baccata	Bark	−−	−−
#114 TBBALM	Penicillium sp	T. baccata	Bark	+	+
#118 TBBALM	Unidentified	T. baccata	Bark	+	+
#120 TBBALM	Pestalotiopsis sp	T. baccata	Bark	+	+
#405 TBBALM	Phomopsis sp	T. baccata	Bark	−−	−−
control(lipase+ RB)
Orlistat				++++	++++

(+) sign indicates the severity of PL inhibition.

Table 2. In vitro porcine pancreatic lipase inhibitory activity of the culture filtrates in phenol red plate assay.

Culture code	Enzyme activity* as halo formed (diameter mm)	% Inhibition
Control	24.4 ± 0.3	0
ORLISTAT	10 ± 0.1	59.0 ± 0.2
#57 TBBALM	16 ± 0.5	34.6 ± 2.5
#1 CSSTOT	16.5 ± 0.4	32.5 ± 1.6
#8 PNLNEY	16.5 ± 0.5	32.5 ± 2.5
#2 CSSTOT	16.7 ± 0.5	31.6 ± 2.4
#33 TBBALM	17.3 ± 0.1	29.2 ± 0.9
#50 TBBALM	17.7 ± 0.2	27.4 ± 0.2
#1 CCSTITD	17.9 ± 0.2	26.8 ± 0.2
#54 TBBALM	18 ± 0.1	26.5 ± 0.2
#7 CSSTOT	18 ± 0.2	26.5 ± 0.2
#2162 CZSTITG	18 ± 0.2	26.4 ± 0.2
#93 TBBALM	19 ± 0.2	22.6 ± 0.3
#7 TBBALM	19 ± 0.1	22.4 ± 0.3
#40 TBBALM	19 ± 0.2	22.1 ± 0.3
#20 TBBALM	19.1 ± 0.3	21.8 ± 0.3
#35 TBBALM	19.2 ± 0.1	21.5 ± 0.3
#31 TBBALM	19.2 ± 0.1	21.5 ± 0.3
#2 PNLNEY	20.3 ± 0.3	16.9 ± 0.5
#14 TBBALM	20.3 ± 0.1	16.8 ± 1.3
#4 TMSDTYEL	21 ± 0.2	14.4 ± 0.5
#16 RSLBRT	21 ± 0.1	14.2 ± 1.3
#11 TBBALM	21.3 ± 0.4	13.0 ± 1.1
#5 CZBAWLS	21.3 ± 0.1	12.9 ± 1.3
#16 NOBASVNP	21.6 ± 0.4	11.6 ± 2.9
#91 TBBALM	22.0 ± 0.1	9.9 ± 2.7
#98 TBBALM	22.1 ± 0.1	9.7 ± 1.3
#89 TBBALM	22.2 ± 0.1	9.2 ± 1.4
#118 TBBALM	22.2 ± 0.1	9.0 ± 1.4
#36 TBBALM	22.3 ± 0.1	8.8 ± 1.5
#42 TBBALM	22.3 ± 0.1	8.7 ± 1.5
#114 TBBALM	22.4 ± 0.1	8.3 ± 1.5
#67 TBBALM	22.6 ± 0.2	7.6 ± 0.6
#46 TBBALM	22.8 ± 0.2	6.8 ± 0.6
#25 TBBALM	23.4 ± 0.2	4.3 ± 0.7
#2106 CZSTITG	23.5 ± 0.1	3.8 ± 0.7
#2112 CZSTITG	23.8 ± 0.4	2.6 ± 0.9
#1 CCBD	23.8 ± 0.4	2.4 ± 0.9
#17 TBBALM	24.0 ± 0.1	1.9 ± 1.6

*Stock concentration of enzyme used 40 U/ml or 0.6 U per well.

Table 3. Inhibitory activity of the endophytic culture filtrates against Pl in rhodamine plate assay.

Culture code	Enzyme activity* as halo formed (diameter mm)	% Inhibition
Control	10 ± 0.7
ORLISTAT	5.2 ± 0.1	48.0 ± 5.3
#57 TBBALM	5.4 ± 0.1	46.1 ± 5.5
#33 TBBALM	5.7 ± 0.0	43.1 ± 8.0
#7 TBBALM	6.4 ± 0.1	35.6 ± 7.0
#35 TBBALM	6.5 ± 0.1	35.2 ± 7.2
#31 TBBALM	6.7 ± 0.2	33.5 ± 5.4
#89 TBBALM	7.0 ± 0.0	30.2 ± 9.8
#91 TBBALM	7.0 ± 0.1	30 ± 7.8
#1 CSSTOT	7.2 ± 0.2	28 ± 6.1
#93TBBALM	7.2 ± 0.2	28 ± 6.1
#20 TBBALM	7.3 ± 0.2	27.4 ± 6.3
#50 TBBALM	7.5 ± 0.5	25.3 ± 0.5
#54 TBBALM	7.7 ± 0.6	23.3 ± 1.2
#8 PNLNEY	7.9 ± 0.4	21.1 ± 1.1
#98 TBBALM	8.2 ± 0.4	18 ± 3.5
#118 TBBALM	8.3 ± 0.0	17.4 ± 11.7
#120 TBBALM	8.4 ± 0.1	16.1 ± 19.8
#67 TBBALM	8.4 ± 0.1	16 ± 19.8
#11 TBBALM	8.4 ± 0.2	16 ± 7.8
#46 TBBALM	8.5 ± 0.4	15 ± 3.9
#2 CSSTOT	8.6 ± 0.3	14.2 ± 6.1
#114 TBBALM	8.6 ± 0.13	14 ± 9.5
#14 TBBALM	8.6 ± 0.2	14 ± 9.1
#16 NOBASVNP	8.6 ± 0.4	14 ± 4.0
#1 CCSTITD	8.6 ± 0.1	13.6 ± 10.0
#42 TBBALM	8.6 ± 0.1	13.6 ± 10.0
#25 TBBALM	8.7 ± 0.3	13.3 ± 6.2
#36 TBBALM	8.8 ± 0.0	12.3 ± 12.4
#2162 CZSTITG	8.7 ± 0.1	11.4 ± 10.2
#5 CZBAWLS	9.0 ± 0.0	10 ± 12.6
#40 TBBALM	9.4 ± 0.0	No activity
#7 CSSTOT	9.4 ± 0.3	No activity
#2 PNLNEY	9.4 ± 0.2	No activity
#4 TMSDTYEL	9.6 ± 0.4	No activity
#16 RSLBRT	9.6 ± 0.3	No activity
#17 TBBALM	9.7 ± 0.4	No activity

*Stock concentration of enzyme used 40 U/ml or 0.6 U per well.

Figure 1. Dose–response curves for inhibition of porcine pancreatic lipase by ethyl acetate extracts of culture filtrates of the selected test fungi. Orlistat is used as a positive control.

Figure 2. Phylogenetic tree showing ITS-based molecular taxonomy and phylogenetic analysis of #57 TBBALM.

Figure 3. Light microscope micrographs of (A): Stipe, metulae and phialides of #57 TBBALM. (B) Conidia. Bar 10 µm.