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  • Research
  • Open Access

Virtual screening for angiotensin I-converting enzyme inhibitory peptides from Phascolosoma esculenta

  • 1,
  • 1,
  • 1,
  • 2,
  • 1Email author and
  • 1
Bioresources and Bioprocessing20141:17

https://doi.org/10.1186/s40643-014-0017-5

©  Liu et al.; licensee Springer. 2014

  • Received: 3 July 2014
  • Accepted: 4 September 2014
  • Published:

Abstract

Background

Many short peptides have proved to exhibit potential anti-hypertensive activity through the inhibition of the Angiotensin I-converting enzyme (ACE) activity and the regulation of blood pressure. However, the traditional experimental screening method for ACE inhibitory peptides is time consuming and costly, accompanied with the limitations as incomplete hydrolysis and peptides loss during purification process. Virtual methods with the aid of computer can break such bottle-neck of experimental work. In this study, an attempt was made to establish a library of di- and tri-peptides derived from proteins of Phascolosoma esculenta, a kind of seafood, through BIOPEP (http://www.uwm.edu.pl/biochemia/index.php/pl/biopep), and to screen highly active ACE inhibitory peptides by molecular docking with the help of LibDock module of Discovery Studio 3.5 software.

Results

Two hundred and eighty four (284) di- and tri-peptides, derived from P. esculenta proteins after a virtual hydrolysis with pepsin, trypsin and a mixture of pepsin and trypsin, were predicted to possess ACE inhibitory activity, among which there are 99 ACE inhibitory peptides with estimated IC50 less than 50 μM. Nine peptides were synthesized for the comparison between the estimated and the experimentally determined IC50. The results indicated that errors between the estimated and measured log(1/IC50) are all less than 1.0 unit.

Conclusions

Virtual method for peptide library construction and ACE inhibitory peptides screening efficiently demonstrated that P. esculenta proteins are prospect resource for food-origin ACE inhibitory peptide.

Keywords

  • Virtual screening
  • Angiotensin I-converting enzyme (ACE)
  • ACE inhibitory peptide
  • Phascolosoma esculenta

1Background

Hypertension is a worldwide health problem, the prevalence of which have affected up to 30% of the adult population according to the World Health Organization. Hypertension carries a high-risk factor for arteriosclerosis, myocardial infarction, and end-stage renal disease [1],[2]. It is predicted that by 2025, about 20% of the world population will suffer from hypertension [3].

Although the cause of hypertension currently cannot be well determined, it is understood that the renin-angiotensin system regulates an organism's water, electrolytes, and blood, and the angiotensin I-converting enzyme (ACE) (peptidyldipeptide hydrolase, EC 3.4.15.1) plays an important role in regulating the blood pressure [4]. ACE is a hypertension-responsible glycoprotein distributed in vascular endothelial, absorptive epithelial, and male germinal cells [5],[6]. ACE cleaves the carboxyl terminal His-Leu dipeptide from inactive decapeptide angiotensin I to active angiotensin II, a powerful vasoconstrictor which can trigger hypertension [7]-[10]. ACE also influences the kallikrein-kinin system by promoting the degradation and inactivation of bradykinin, which can lead to reduction of hypertension. Therefore, excessive activity of ACE leads to hypertension. Molecules which can inhibit the activity of ACE are considered useful drugs for hypertension management [11]. Currently, synthetic ACE inhibitors, such as captopril, enalapril and lisinopril, are available on the market [12]; however, they tend to have side effects [13].

Since the discovery of the first anti-hypertensive peptide in snake venom [14], more attention has been paid to natural sources, especially peptides. Peptides derived from cheese whey [15], fermented milk [16], mushroom [17], soy bean [18],[19], corn gluten [20], insect protein [21], peanut flour [22], and egg [23] have been proven to inhibit the activity of ACE. However, few studies were reported about their side effects [24],[25]. Nutritionists claim that peptides found in food are safer than ‘traditional’ drugs, and they are promising synthetic drug substitutes [26].

Among the ACE inhibitory peptides, shorter ones (di- and tri-peptides) usually have significant advantages over longer ones. They easily pass through blood circulation system [27],[28] and then reach action sites faster without being hydrolyzed by digestive enzymes during the gastrointestinal digestion [29],[30]. For these reasons, the present study focused on di- and tri-peptides.

The discovery of ACE inhibitory peptides with potential anti-hypertensive effect is mostly based on experiments, which require amounts of labors and funds. Besides, the possible active peptides can not be totally harvested due to the incomplete hydrolysis and peptides loss in the purification by the experimental protocols. Recently, as the computation simulation technology for drug design and discovery of molecular interaction are booming, the virtual screening or in silico experiment may replace the traditionally experimental screening of anti-hypertensive peptides to some extent. Computational approaches, which are based on computational evaluation of interactions between receptor and ligand, are proved feasible for virtual screening [31]. Molecular docking is a powerful and a widely used tool in molecular simulation, which is approximated to a lock-and-key process. The docking protocol is to ‘dock’ a ligand into an active site of a receptor; then, the interactions between them were ‘scored’ to assess the potential bioactivity of candidate compounds. The most advantage of docking is its high-throughput screening in short time with little cost [32].

In this study, an attempt was made to investigate the ACE inhibitory activity of di- and tri-peptides derived from Phascolosoma esculenta, a marine deposit-feeding benthonic invertebrates, also a traditional seafood with over 70% protein (dry weight) in Southeast China [33],[34]. Database of di- and tri-peptides derived from P. esculenta were established, and their ACE inhibitory activities were predicted by virtual hydrolysis and screening method. Finally, di- and tri-peptides which have obvious ACE inhibitory activity were synthesized for verifying the validity of such virtual strategy.

2Methods

2.1 Materials

There are 22 proteins of P. esculenta with the protein messages including entry name and sequence obtained from UniProt (http://www.uniprot.org/) (Table 1). They were used as original materials for database of di- and tri-peptides. With the help of BIOPEP (http://www.uwm.edu.pl/biochemia/index.php/pl/biopep), the 22 proteins were virtually hydrolyzed with pepsin, trypsin, and a mixture of pepsin and trypsin.
Table 1

Properties of 22 P. esculenta proteins in UniProt ( http://www.uniprot.org/ )

Name

Number of amino acids

MW (kDa)

PI

D2J0B2

226

25.2

6.0

C3PUI4

378

43.2

9.1

D2J288

726

83.6

4.9

C3PUI8

304

34.6

8.5

C3PUI2

267

30.3

6.9

B6CQR3

658

71.6

5.1

A5A2J9

135

15.4

6.5

C3PUI5

121

13.9

6.7

B3TCX0

84

9.3

4.7

C3PUJ0

571

63.3

9.2

C3PUI9

231

25.7

5.9

B6CPA3

120

13.63

5.8

A5A2K2

220

24.4

5.8

C3PUH9

519

57.2

6.2

C3PUI7

450

50.2

9.2

B3TFG2

174

20.2

5.1

C3PUI1

54

6.5

10.8

C3PUI3

157

17.7

9.5

A3EX91

137

14.8

4.7

C3PUJ1

323

36.1

9.1

C3PUI0

231

25.9

4.8

C3PUI6

94

10.5

9.2

MW molecular weight, PI isoelectric point.

2.2 Molecular docking experiments

LibDock, a module of Discovery Studio 3.5 software (DS3.5, Accelrys, San Diego, CA, USA), was used for molecular docking experiments. Scoring results (LibDock score) about ligand-receptor combination were used as the final criterion to estimate the ACE inhibitory activity of ligands. Based on a previous study [35], the corresponding relationship between LibDock score and IC50 was
LibDock score = 10.063 log 1 / IC 50 + 68.08
,

where IC50 is 50% inhibitory concentration (in μM) towards ACE. According to the LibDock score, ACE inhibitory activity of ligands could be estimated.

ACE was used as receptor in docking simulation, whose crystal structures was available in the Protein Data Bank (PDB) (http://www.pdb.org), from where the three-dimensional structure of ACE was imported [PDB:1O8A]. Before the docking procedure, water molecules were removed and zinc ions were retained. The 284 di- and tri-peptides derived from P. esculenta were used as ligands, of which structures and energies were generated with ChemBioDraw software [36] and minimized with the CHARMM program [37], respectively. Parameters used in the docking process are shown in Table 2.
Table 2

Parameters for molecular docking experiments performed with the LibDock of DS3.5

Parameter name

Parameter value

Docking sphere

10 Å

Input site sphere

 

 x

48.65

 y

82.55

 z

54.04

Number of HPTPot

100

Docking tolerance

0.25

Docking preference

User specified

Max hits to save

10

Max number of hits

100

Minimum LibDock score

100

Final score cutoff

0.5

Max BFGS steps

50

Rigid optimization

False

Max conformation hits

30

Max start conformations

1,000

Steric fraction

0.10

Final cluster radius

0.5

Apolar SASA cutoff

15.0

Polar SASA cutoff

5.0

Surface grid steps

18

Conformation method

Best

Minimization algorithm

Do not minimize

Parallel processing

True

2.3 Synthesis of peptides

Five tri-peptides (GYF, WAL, AYF, GLR, and ILK) and four di-peptides (FK, QF, EL, and HK) generated through in silico hydrolysis of P. esculenta protein, with purity of 95%, were synthesized by GL Biochem Co. Ltd. (Shanghai, China) for IC50 testing.

2.4 Measurement of ACE inhibitory activity

The ACE inhibitory activity was measured according to the method of Cushman and Cheung [38] with slight modifications. Ten milligram of the sample was dissolved in 1 mL distilled water and then diluted to seven different concentrations for ACE inhibitory measurements. Fifteen microliters of the sample solution in certain concentration (Seven different concentrations) were needed, which were determined by the pre-experiment about ACE inhibition ratio. The whole principle is that the concentration which ACE inhibition ratio reaches 50% is included within the concentration range. The concentrations for GYF, FK, WAL, QF, and AYF are 10, 20, 30, 40, 50, 60, and 70 μg/mL, and for EL, GLR, HK, and ILK are 20, 40, 60, 80, 100, 120, and 140 μg/mL, respectively) and 15 μL substrate hippuryl-l-histidyl-l-leucine (HHL) (8.3 mM Hip-His-Leu in 50 mM sodium borate buffer containing 0.5 M NaCl at pH 8.3) were mixed together and then pre-incubated at 37°C for 5 min. The reaction was initiated by adding 5 μL of ACE solution (310 mU/mL) and incubated for 60 min at the same temperature. The reaction was terminated by the addition of 1.0 M HCl (200 μL). Ten microliters of the reaction solution was injected directly onto a Thermo BDS-C18 column (3.0 mm × 250 mm, 5 μm, Thermo Scientific Co. Ltd., Waltham, MA, USA). The mobile phase consisted of 10% acetonitrile and 90% water with 0.1% trifluoroacetic acid (TFA). The flow rate was 0.7 mL/min and the absorbance was monitored at 228 nm. All determination was carried out at least in triplicate. The inhibition activity was calculated using the following equation:
ACE inhibition % = 1 − A inhibitor / A control × 100
,

where Ainhibitor is the absorbance with ACE, HHL, and sample, and Acontrol is the absorbance of hippuric acid (HA) with ACE and HHL without the sample. Dose-dependent ACE inhibition was investigated using at least five different concentrations of peptides. The concentration of peptides that inhibited ACE activity by 50% (IC50) was calculated using a non-linear regression from a plot of ACE inhibition versus sample concentrations.

2.5 Study on structural-active relationship of ACE inhibitory peptides

The chemical properties of C-terminal and N-terminal amino acids of 99 peptides with estimated IC50 less than 50 μM were summarized to deduce the structural-active relationship of ACE inhibitory peptides.

3Results and discussion

3.1 Pool of di- and tri-peptides derived from P. esculenta proteins

Pepsin, trypsin, and the mixture of pepsin and trypsin were used to virtually hydrolyze the 22 proteins from P. esculenta, with the help of BIOPEP (http://www.uwm.edu.pl/biochemia/index.php/pl/biopep). In total, 2,667 peptides were virtually produced, and among them, 1,084 were di- and tri-peptides, which accounted for about 40.6% (Figure 1). After excluding the repeated ones, there were 1,017 non-repeated peptides, among which 284 were di- and tri-peptides. The sequences and the frequencies of these 284 short peptides are shown in Table 3. These 284 peptides were used as the ligands for docking experiment with ACE.
Figure 1
Figure 1

Length distribution of peptides derived from P. esculenta proteins by virtual hydrolysis. The enzymes used in virtual hydrolysis are pepsin, trypsin, and a mixture of pepsin and trypsin.

Table 3

Sequence and frequency of di- and tri-peptides derived from P. esculenta proteins by virtual hydrolysis

Peptide

Frequency

Peptide

Frequency

Peptide

Frequency

Peptide

Frequency

ADL

1

GYF

2

NDK

2

STL

2

AEF

2

HAQ

2

NF

10

SVF

2

AEK

1

HER

1

NGK

1

SVL

4

AEL

2

HF

9

NIL

2

SWK

2

AF

19

HGL

2

NK

2

SWL

2

AGF

2

HIK

2

NL

13

SYL

1

AIF

4

HK

4

NLK

1

TAL

2

AK

2

HKF

1

NMR

1

TDK

1

AL

35

HL

17

NPF

6

TDR

1

ALR

1

HSL

2

NR

2

TEF

2

AMF

2

HTK

1

NRF

2

TF

7

APF

2

HTL

1

NSF

2

TGF

2

AQF

2

IAR

2

NTL

3

TGL

4

AR

2

ICL

4

NVL

5

TIL

2

ASK

1

IF

20

NVR

2

TK

4

AVK

2

IGR

1

NWL

2

TL

31

AVR

2

IHR

2

PCK

1

TLK

1

AYA

2

IIL

4

PDL

2

TML

2

AYF

3

IK

7

PF

19

TNR

1

CF

2

IL

46

PGF

2

TPF

1

CK

1

ILK

1

PIL

2

TSL

4

CL

9

IMF

2

PK

7

TTK

1

CVF

2

IMK

2

PKL

1

TVK

1

DF

4

IPK

2

PL

30

TVL

2

DK

16

IPL

5

PNK

2

TVR

1

DL

11

IQK

2

PPL

2

TWK

1

DMF

2

IR

4

PRL

1

TYF

2

DNR

1

IRF

1

PSF

2

VAL

6

DPK

1

ISF

2

PSK

1

VDL

2

DR

4

ISL

2

PSL

2

VEK

3

DSK

2

ISR

1

PTL

4

VER

2

DSL

4

ITK

1

PTR

2

VF

9

DWL

1

ITL

2

PVK

1

VGF

6

EAF

2

IVL

1

PVL

2

VGL

3

EAR

1

IWL

4

QAL

2

VIR

3

EDK

2

KAL

1

QDF

2

VK

2

EEF

1

KDL

2

QEL

2

VKL

1

EEL

2

KF

2

QF

5

VL

14

EER

1

KGF

1

QGL

2

VMK

2

EF

2

KIL

1

QIR

1

VML

2

EGL

1

KL

4

QK

2

VNL

4

EIF

1

KPL

1

QL

7

VPK

1

EIL

1

KRF

1

QR

1

VPL

4

EK

7

KSL

1

QYK

1

VSF

2

EL

16

KTL

1

RAL

1

VSL

3

EML

1

KVF

1

REL

1

VVF

2

ENK

2

LDK

1

RL

3

VVK

2

ENL

6

LEK

1

RPF

1

WAF

2

ER

2

LK

2

RSF

2

WAL

2

ESK

1

LMK

1

RTL

1

WCF

2

ETL

3

LR

1

SAL

4

WF

6

EVK

1

LSK

1

SEK

1

WGK

2

EVL

4

MAL

8

SF

20

WL

8

FFK

1

MF

16

SGF

4

WML

2

FK

1

MFK

1

SGL

2

WNF

2

FWR

1

MFR

1

SHL

2

WPF

2

GAL

2

MGF

2

SIF

2

WQK

2

GF

12

MGL

2

SIK

2

WR

1

GGL

4

MIK

2

SIL

4

WTR

1

GGR

2

MIL

2

SK

6

WWF

2

GK

6

MK

10

SL

50

YAL

1

GKF

1

MKF

2

SNL

3

YF

7

GL

29

ML

16

SPF

2

YIF

2

GLR

1

MPL

1

SPL

2

YIK

1

GNL

2

MR

4

SQL

2

YK

4

GR

3

MSK

2

SR

2

YL

8

GSL

2

MSL

10

SS

2

YPL

2

GTL

4

MTK

2

SSF

6

YS

2

GTR

2

MTL

2

SSL

2

YSK

3

GVK

1

MVK

2

STF

2

YTL

2

GWL

2

NAL

2

STK

1

YVR

1

The enzymes used in virtual hydrolysis are pepsin, trypsin, and a mixture of pepsin and trypsin.

3.2 Estimated IC50 distribution of ACE inhibitory di- and tri-peptides

The estimated ACE inhibitory IC50 of the 284 di- and tri-peptides derived from P. esculenta proteins were obtained according to LibDock scores, which were summarized in Figure 2. Ninety-nine (99) peptides had an estimated IC50 less than 50 μM (34.9% of 284 peptides), 100 peptides had an estimated IC50 between 50 and 100 μM (35.2% of 284 peptides), and 37 peptides had an estimated IC50 less than 500 μM, accounting for 13.0%. Most reported ACE inhibitory peptides with IC50 less than 100 μM showed potent in vivo anti-hypertensive activity [39]. Therefore, P. esculenta is a prospective anti-hypertensive peptide-containing resource since more than two thirds di- and tri-peptides theoretically possess obvious ACE inhibitory activity. The sequences and estimated IC50 of di- and tri-peptides with estimated IC50 less than 50 μM are shown in Table 4.
Figure 2
Figure 2

Distribution of estimated IC 50 of di- and tri-peptides with ACE inhibitory activity. The peptides were derived from P. esculenta proteins. All IC50 values were predicted by LibDock scores according to the equation, LibDock score = 10.063 log(1/IC50) + 68.08.

Table 4

Di- and tri-peptides derived from P. esculenta with estimated IC 50 less than 50 μM

Peptide

Estimated IC50

Peptide

Estimated IC50

Peptide

Estimated IC50

WNF

0.12

YIF

13.1

DSL

27.9

HKF

0.42

RPF

13.3

IR

28.9

WCF

0.47

APF

13.4

TGF

29.5

WPF

0.85

REL

13.7

NWL

30.7

WWF

1.18

YK

13.9

FFK

30.9

YVR

1.45

AEF

14.1

GYF

32.2

NRF

1.71

AYF

14.7

ISF

32.3

TYF

1.75

KTL

14.8

FK

33.2

IHR

2.06

WGK

14.9

SPF

33.4

WML

2.15

DWL

14.9

VML

33.7

WAF

3.27

SSF

15.0

KPL

36.1

IWL

5.22

NSF

15.4

ISL

36.5

RSF

5.44

VKL

16.1

QF

36.7

IRF

5.72

MAL

16.3

TML

36.9

CVF

7.15

NR

17.0

QEL

37.0

YIK

7.33

MTL

17.1

MKF

38.0

MGF

7.38

IMF

17.5

YAL

38.6

YF

7.48

WR

17.7

EEF

40.4

KRF

8.11

ER

18.1

IGR

40.5

SR

9.05

AEL

19.4

TEF

41.1

GNL

9.09

QDF

19.5

TTK

41.6

GKF

9.51

YTL

19.7

SGF

41.6

WTR

10.2

PIL

19.9

KVF

42.0

PRL

10.2

KDL

20.2

SIL

43.0

SYL

10.4

SVF

20.8

EVL

43.7

WF

10.6

YSK

21.5

DMF

46.6

IIL

10.8

VVF

22.5

RTL

46.9

WAL

11.3

ITK

22.6

QAL

47.0

AQF

11.5

DR

23.4

TPF

47.0

RAL

11.8

WQK

23.9

HSL

47.1

PGF

12.2

PKL

24.9

NAL

47.1

NVL

12.7

EIF

25.2

SHL

48.0

HTL

13.1

VGF

27.8

STF

49.4

Short peptides were usually used for predicting potent ACE inhibitory activity. Pripp docked 58 di-peptides into protein target using the Molegro Virtual Docker version 1.1.1 software and found significant relationship between docking results and experimental IC50 values [32]. Several tri-peptides consisting of I or L and positive charged amino acids and aromatic amino acids were synthesized, and their ACE inhibitory activities were measured to clarify the amino acid sequence for inhibition of ACE [40]. Larger peptides, for instance, the sequence length more than 5, were also focused in some work [41]; however, such works were reported with lower R2 (coefficient of variation) because of the complexity in the modeling due to the bigger peptide [42],[43].

3.3 Confirmation of virtual screening method

In order to confirm the validity of virtual screening method of the present work, nine peptides were synthesized for IC50 testing. The sequences of these peptides were obtained from P. esculenta protein through virtual hydrolysis and screening by docking experiments. The estimated log(1/IC50) and measured log(1/IC50) of the nine peptides were compared (Table 5). The error between the estimated log(1/IC50) and measured log(1/IC50) is less than 1.0 unit. Desirable limit for model is that the error between estimated log(1/IC50) and measured log(1/IC50) is less than 1.5 units [30]. A reported quantitative structure-activity relationship (QSAR) model was constructed on 168 di-peptides and 140 tri-peptides collected from literatures, and the model verification was made on seven reported di-peptides and tri-peptides (not included in 168 di-peptides and 140 tri-peptides), of which the error was between 0.07 and 1.39 [29]. On the ground of such criterion, the present model is efficient and credible.
Table 5

Estimated and measured log(1/IC 50 ) of the nine synthesized peptides derived from P. esculenta

Peptide

Estimated log(1/IC50)

Measured log(1/IC50)

Error

GYF

4.49

4.31 ± 0.02

0.18

FK

4.79

4.27 ± 0.01

0.52

WAL

4.95

4.38 ± 0.04

0.57

QF

4.44

4.21 ± 0.03

0.23

AYF

4.83

4.18 ± 0.01

0.65

EL

3.19

3.43 ± 0.02

−0.24

GLR

4.07

3.61 ± 0.02

0.46

HK

3.53

3.86 ± 0.01

−0.33

ILK

4.09

3.72 ± 0.03

0.37

Previous studies suggested that the structural-active relationship of ACE inhibitory peptides largely depended on their amino acid composition, sequence, and configuration, though the full mechanism of interaction between peptides and ACE is not established so far [44],[45]. For the short peptides as di- and tri-peptides, the amino acid composition and configuration are more significant. The di- and tri-peptides which have an estimated IC50 within 50 μM were used to study the structural-active relationship of these ACE inhibitors.

There are four kinds of C-terminal residues for 99 sequences (Figure 3) due to the cutting specificity of pepsin and trypsin. Leu and Phe are C-terminal residues formed by pepsin hydrolysis, and C-terminal Lys and Arg are formed by trypsin reaction. Hydrophobic C-terminal (Phe and Leu) is dominant in amount and accounts for more than 80% peptides (44.4% and 36.4%, respectively). There are some accepted concepts about the structure-activity relationship of ACE inhibitory peptides, such as that peptides with hydrophobic amino acid in C-terminus showed a highly potent ACE inhibitory activity [46]. Highly active peptide in general should be composed of large, hydrophobic, and aromatic amino acid with a polar functional group in C-terminus [47]; and the physicochemical attributes of amino acids such as hydrophobicity, bulkiness, and electronic properties had impacts on the bioactivity of peptides [48]. Accordingly, benzene ring in Phe can also increase the bulkiness and bring about the stability of binding between ACE and peptide and sequentially result in high ACE inhibitory activity.
Figure 3
Figure 3

Proportions of C-terminal amino acids for di- and tri-peptides with estimated IC 50 less than 50 μM.

There are 40 peptides among 99 peptides (40.4%) with hydrophobic amino acid at N-terminal, 38 peptides with neutral amino acid at N-terminal (38.4%), and 21.9% peptides with positively or negatively charged amino acid at N-terminal (Figure 4). N-terminal amino acid of ACE inhibitory peptides also favors the hydrophobic interactions with ACE [7],[30]. The peptides with hydrophobic amino acid at N-terminal showing higher ACE inhibitory activity have some superiority in amount in the present study, which verified such view.
Figure 4
Figure 4

Proportions of N-terminal amino acids for di- and tri-peptides with estimated IC 50 less than 50 μM.

4Conclusions

A virtual method of hydrolysis and screening of ACE inhibitory peptides with high activity such as IC50 value < 50 μM was constructed in this work. Ninety-nine (99) peptides were obtained from 22 proteins of P. esculenta. Besides, the efficiency and the validity of such method were verified by comparing the predicted IC50 and measured IC50 of some synthesized peptides among the 99 peptides. The results demonstrated that the virtual hydrolysis and screening method is an efficient way that greatly cuts down the experimental labor to get highly active ACE inhibitory peptides. Moreover, P. esculenta proteins were proved as a good resource of ACE inhibitory peptides, which could be a beneficial ingredient for functional foods or pharmaceuticals against hypertension. Further research on larger anti-hypertension peptides derived from P. esculenta and in vivo activity testing will be carried out.

Declarations

Acknowledgements

This work was supported by ‘National Natural Science Foundation of China (No. 31301413)’, ‘National Major Science and Technology Projects of China (No. 2012ZX09304009)’, and the ‘Fundamental Research Funds for the Central Universities’, People's Republic of China.

Authors’ Affiliations

(1)
State Key Laboratory of Bioreactor Engineering, Department of Food Science and Engineering, East China University of Science and Technology, 130# Meilong Rd., Shanghai, 200237, People’s Republic of China
(2)
Zhejiang Key Lab of Exploitation and Preservation of Coastal Bio-resource, Wenzhou, 325005, People’s Republic of China

References

  1. Jung WK, Mendis E, Je JY, Park PJ, Son BW, Kim HC, Choi YK, Kim SK: Angiotensin I-converting enzyme inhibitory peptide from yellowfin sole ( Limanda aspera ) frame protein and its antihypertensive effect in spontaneously hypertensive rats. Food Chem 2006,94(1):26–32. 10.1016/j.foodchem.2004.09.048View ArticleGoogle Scholar
  2. Silva DG, Freitas MP, da Cunha EFF, Ramalho TC, Nunes CA: Rational design of small modified peptides as ACE inhibitors. Med Chem Comm 2012,3(10):1290–1293. 10.1039/c2md20214jView ArticleGoogle Scholar
  3. Kearney PM, Whelton M, Reynolds K, Muntner P, Whelton PK, He J: Global burden of hypertension: analysis of worldwide data. Lancet 2005,365(9455):217–223. 10.1016/S0140-6736(05)17741-1View ArticleGoogle Scholar
  4. Li GH, Le GW, Shi YH, Shrestha S: Angiotensin I-converting enzyme inhibitory peptides derived from food proteins and their physiological and pharmacological effects. Nutr Res 2004,24(7):469–486. 10.1016/j.nutres.2003.10.014View ArticleGoogle Scholar
  5. Guang C, Phillips RD: Plant food-derived angiotensin I converting enzyme inhibitory peptides. J Agric Food Chem 2009,57(12):5113–5120. 10.1021/jf900494dView ArticleGoogle Scholar
  6. De Leo F, Panarese S, Gallerani R, Ceci L: Angiotensin converting enzyme (ACE) inhibitory peptides: production and implementation of functional food. Curr Pharm Des 2009,15(31):3622–3643. 10.2174/138161209789271834View ArticleGoogle Scholar
  7. Iroyukifujita H, Eiichiyokoyama K, Yoshikawa M: Classification and antihypertensive activity of angiotensin I-converting enzyme inhibitory peptides derived from food proteins. J Food Sci 2000,65(4):564–569. 10.1111/j.1365-2621.2000.tb16049.xView ArticleGoogle Scholar
  8. Reneland R, Lithell H: Angiotensin-converting enzyme in human skeletal muscle. A simple in vitro assay of activity in needle biopsy specimens. Scand J Clin Lab Investig 1994,54(2):105–111. 10.3109/00365519409086516View ArticleGoogle Scholar
  9. Hartl FU: Molecular chaperones in cellular protein folding. Nature 1996, 381: 571–580. 10.1038/381571a0View ArticleGoogle Scholar
  10. Sturrock E, Natesh R, Van Rooyen J, Acharya K: Structure of angiotensin I-converting enzyme. Cell Mol Life Sci 2004, 61: 2677–2686. 10.1007/s00018-004-4239-0View ArticleGoogle Scholar
  11. Lin L, Lv S, Li B: Angiotensin-I-converting enzyme (ACE)-inhibitory and antihypertensive properties of squid skin gelatin hydrolysates. Food Chem 2012,131(1):225–230. 10.1016/j.foodchem.2011.08.064View ArticleGoogle Scholar
  12. Sweitzer NK: What is an angiotensin converting enzyme inhibitor? Circulation 2003,108(3):e16-e18. 10.1161/01.CIR.0000075957.16003.07View ArticleGoogle Scholar
  13. Antonios TF, MacGregor GA: Angiotensin converting enzyme inhibitors in hypertension: potential problems. J Hypertens 1995, 13: S11-S16. 10.1097/00004872-199509003-00003View ArticleGoogle Scholar
  14. Ondetti MA, Williams NJ, Sabo E, Pluscec J, Weaver ER, Kocy O: Angiotensin-converting enzyme inhibitors from the venom of Bothrops jararaca . Isolation, elucidation of structure, and synthesis. Biochemistry 1971,10(22):4033–4039. 10.1021/bi00798a004View ArticleGoogle Scholar
  15. Abubakar A, Saito T, Kitazawa H, Kawai Y, Itoh T: Structural analysis of new antihypertensive peptides derived from cheese whey protein by proteinase K digestion. J Dairy Sci 1998,81(12):3131–3138. 10.3168/jds.S0022-0302(98)75878-3View ArticleGoogle Scholar
  16. Nakamura Y, Yamamoto N, Sakai K, Okubo A, Yamazaki S, Takano T: Purification and characterization of angiotensin I-converting enzyme inhibitors from sour milk. J Dairy Sci 1995,78(4):777–783. 10.3168/jds.S0022-0302(95)76689-9View ArticleGoogle Scholar
  17. Andújar-Sánchez M, Cámara-Artigas A, Jara-Pérez V: A calorimetric study of the binding of lisinopril, enalaprilat and captopril to angiotensin-converting enzyme. Biophys Chem 2004,111(2):183–189. 10.1016/j.bpc.2004.05.011View ArticleGoogle Scholar
  18. Wu J, Ding X: Hypotensive and physiological effect of angiotensin converting enzyme inhibitory peptides derived from soy protein on spontaneously hypertensive rats. J Agric Food Chem 2001,49(1):501–506. 10.1021/jf000695nView ArticleGoogle Scholar
  19. Mallikarjun Gouda K, Gowda LR, Rao AA, Prakash V: Angiotensin I-converting enzyme inhibitory peptide derived from glycinin, the 11S globulin of soybean (Glycine max). J Agric Food Chem 2006,54(13):4568–4573. 10.1021/jf060264qView ArticleGoogle Scholar
  20. Suh H, Whang J, Lee H: A peptide from corn gluten hydrolysate that is inhibitory toward angiotensin I converting enzyme. Biotechnol Lett 1999,21(12):1055–1058. 10.1023/A:1005688627350View ArticleGoogle Scholar
  21. Vercruysse L, Van Camp J, Morel N, Rougé P, Herregods G, Smagghe G: Ala-Val-Phe and Val-Phe: ACE inhibitory peptides derived from insect protein with antihypertensive activity in spontaneously hypertensive rats. Peptides 2010,31(3):482–488. 10.1016/j.peptides.2009.05.029View ArticleGoogle Scholar
  22. Quist EE, Phillips RD, Saalia FK: Angiotensin converting enzyme inhibitory activity of proteolytic digests of peanut ( Arachis hypogaea L.) flour. LWT-Food Sci Technol 2009,42(3):694–699. 10.1016/j.lwt.2008.10.008View ArticleGoogle Scholar
  23. Majumder K, Wu J: Angiotensin I converting enzyme inhibitory peptides from simulated in vitro gastrointestinal digestion of cooked eggs. J Agric Food Chem 2009,57(2):471–477. 10.1021/jf8028557View ArticleGoogle Scholar
  24. Wang C, Tian J, Wang Q: ACE inhibitory and antihypertensive properties of apricot almond meal hydrolysate. Eur Food Res Technol 2011,232(3):549–556. 10.1007/s00217-010-1411-7View ArticleGoogle Scholar
  25. Vermeirssen V, Camp JV, Verstraete W: Bioavailability of angiotensin I converting enzyme inhibitory peptides. Br J Nutr 2004,92(03):357–366. 10.1079/BJN20041189View ArticleGoogle Scholar
  26. Jimsheena V, Gowda LR: Angiotensin I-converting enzyme (ACE) inhibitory peptides derived from arachin by simulated gastric digestion. Food Chem 2011,125(2):561–569. 10.1016/j.foodchem.2010.09.048View ArticleGoogle Scholar
  27. Seppo L, Jauhiainen T, Poussa T, Korpela R: A fermented milk high in bioactive peptides has a blood pressure-lowering effect in hypertensive subjects. Am J Clin Nutri 2003,77(2):326–330.Google Scholar
  28. Mathews D, Adibi S: Peptide absorption. Gastroenterology 1976,71(1):151.Google Scholar
  29. Wu J, Aluko RE, Nakai S: Structural requirements of angiotensin I-converting enzyme inhibitory peptides: quantitative structure-activity relationship study of di-and tripeptides. J Agric Food Chem 2006,54(3):732–738. 10.1021/jf051263lView ArticleGoogle Scholar
  30. Wijesekara I, Qian ZJ, Ryu B, Ngo DH, Kim SK: Purification and identification of antihypertensive peptides from seaweed pipefish ( Syngnathus schlegeli ) muscle protein hydrolysate. Food Res Int 2011,44(3):703–707. 10.1016/j.foodres.2010.12.022View ArticleGoogle Scholar
  31. Berman HM, Westbrook J, Feng Z, Gilliland G, Bhat T, Weissig H, Shindyalov IN, Bourne PE: The Protein Data Bank. Nucleic Acids Res 2000,28(1):235–242. 10.1093/nar/28.1.235View ArticleGoogle Scholar
  32. Pripp AH: Docking and virtual screening of ACE inhibitory dipeptides. Eur Food Res Technol 2007,225(3–4):589–592. 10.1007/s00217-006-0450-6View ArticleGoogle Scholar
  33. Su X, Du L, Li Y, Li T, Li D, Wang M, He J: Production of recombinant protein and polyclonal mouse antiserum for ferritin from Sipuncula Phascolosoma esculenta . Fish Shellfish Immunol 2009,27(3):466–468. 10.1016/j.fsi.2009.06.014View ArticleGoogle Scholar
  34. Du L, Fang M, Wu H, Xie J, Wu Y, Li P, Zhang D, Huang Z, Xia Y, Zhou L: A novel angiotensin I-converting enzyme inhibitory peptide from Phascolosoma esculenta water-soluble protein hydrolysate. J Funct Foods 2013,5(1):475–483. 10.1016/j.jff.2012.12.003View ArticleGoogle Scholar
  35. Wu H, Liu Y, Guo M, Xie J, Jiang X: A virtual screening method for inhibitory peptides of angiotensin I converting enzyme. J Food Sci 2014, 79: C1635-C1642. doi:10.1111/1750–3841.12559 doi:10.1111/1750-3841.12559 10.1111/1750-3841.12559View ArticleGoogle Scholar
  36. Kerwin SM: ChemBioOffice Ultra 2010 suite. J Am Chem Soc 2010,132(7):2466–2467. 10.1021/ja1005306View ArticleGoogle Scholar
  37. Brooks BR, Bruccoleri RE, Olafson BD, States DJ, Swaminathan S, Karplus M: CHARMM: a program for macromolecular energy, minimization, and dynamics calculations. J Comput Chem 1983,4(2):187–217. 10.1002/jcc.540040211View ArticleGoogle Scholar
  38. Cushman D, Cheung H: Spectrophotometric assay and properties of the angiotensin-converting enzyme of rabbit lung. Biochem Pharmacol 1971,20(7):1637–1648. 10.1016/0006-2952(71)90292-9View ArticleGoogle Scholar
  39. Iwaniak A, Minkiewicz P, Darewicz M: Food-originating ACE inhibitors, including antihypertensive peptides, as preventive food components in blood pressure reduction. Compr Rev Food Sci Food Safety 2014,13(2):114–134. 10.1111/1541-4337.12051View ArticleGoogle Scholar
  40. Kobayashi Y, Yamauchi T, Katsuda T, Yamaji H, Katoh S: Angiotensin-I converting enzyme (ACE) inhibitory mechanism of tripeptides containing aromatic residues. J Biosci Bioeng 2008,106(3):310–312. 10.1263/jbb.106.310View ArticleGoogle Scholar
  41. Sagardia I, Roa-Ureta RH, Bald C: A new QSAR model, for angiotensin I-converting enzyme inhibitory oligopeptides. Food Chem 2013,136(3):1370–1376. 10.1016/j.foodchem.2012.09.092View ArticleGoogle Scholar
  42. Wu J, Aluko RE, Nakai S: Structural requirements of angiotensin I-converting enzyme inhibitory peptides: quantitative structure-activity relationship modeling of peptides containing 4–10 amino acid residues. QSAR Combinat Sci 2006,25(10):873–880. 10.1002/qsar.200630005View ArticleGoogle Scholar
  43. Pripp AH, Isaksson T, Stepaniak L, Søhaug T: Quantitative structure-activity relationship modelling of ACE-inhibitory peptides derived from milk proteins. Eur Food Res Technol 2004,219(6):579–583. 10.1007/s00217-004-1004-4View ArticleGoogle Scholar
  44. Kim SY, Je JY, Kim SK: Purification and characterization of antioxidant peptide from hoki ( Johnius belengerii ) frame protein by gastrointestinal digestion. J Nutr Biochem 2007,18(1):31–38. 10.1016/j.jnutbio.2006.02.006View ArticleGoogle Scholar
  45. Ruiz-Giménez P, Marcos JF, Torregrosa G, Lahoz A, Fernández-Musoles R, Valles S, Alborch E, Manzanares P, Salom JB: Novel antihypertensive hexa- and heptapeptides with ACE-inhibiting properties: from the in vitro ACE assay to the spontaneously hypertensive rat. Peptides 2011,32(7):1431–1438. 10.1016/j.peptides.2011.05.013View ArticleGoogle Scholar
  46. Kapel R, Rahhou E, Lecouturier D, Guillochon D, Dhulster P: Characterization of an antihypertensive peptide from an Alfalfa white protein hydrolysate produced by a continuous enzymatic membrane reactor. Process Biochem 2006,41(9):1961–1966. 10.1016/j.procbio.2006.04.019View ArticleGoogle Scholar
  47. Pripp AH: Initial proteolysis of milk proteins and its effect on formation of ACE-inhibitory peptides during gastrointestinal proteolysis: a bioinformatic, in silico , approach. Eur Food Res Technol 2005,221(5):712–716. 10.1007/s00217-005-0083-1View ArticleGoogle Scholar
  48. Wu J, Aluko RE: Quantitative structure-activity relationship study of bitter di-and tri-peptides including relationship with angiotensin I-converting enzyme inhibitory activity. J Pept Sci 2007,13(1):63–69. 10.1002/psc.800View ArticleGoogle Scholar

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© Liu et al.; licensee Springer. 2014

This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited.

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