Professional Documents
Culture Documents
Review
School of Biosystems and Food Engineering, University College Dublin, Beleld, Dublin 4, Ireland
Department of Food Biosciences, Teagasc Food Research Centre, Dublin 15, Ireland
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 3 January 2015
Received in revised form
23 June 2015
Accepted 14 July 2015
Available online 17 July 2015
Bioactive peptides in foods can be obtained from plants, animal or marine sources. Bioactive peptides
possess diverse biological activities such as opiate, antithrombotic, anti-hypertensive, immunomodulating, antilipemic, osteoprotective, antioxidative, antimicrobial, ileum contracting, anticariogenic and
growth promoting properties. In general, peptide based drugs are physically and chemically unstable,
have short in vivo half-lives and have low oral bioavailability. Ultrasound which is a novel, robust, green
and rapid technology suitable for scale up, can enhance the efciency of protein digestion, extraction,
production and drug delivery of bioactive peptides. Ultrasound principally acts by generating bubble
cavitation in the biological matrix. It has been extensively reported for extraction of proteins and peptides from natural products facilitating higher yields and rates of extraction. Ultrasound assisted
encapsulation of peptide based drugs with biodegradable polymers can improve stability and bioavailability. Moreover, in sonophoresis applications, low-frequency ultrasound can be used to transport high
molecular weight peptide drugs. This review paper summarizes key bioactive peptides, sources, structure, function and application of ultrasound for production and drug delivery of peptides.
2015 Elsevier Ltd. All rights reserved.
Keywords:
Ultrasound
Bioactive peptide
Digestion
Extraction
Drug delivery
Microencapsulation
ACE inhibitory peptide
1. Introduction
Recent developments in functional food research have
generated renewed interest in bioactive compounds including
bioactive peptides. The word peptide comes from the Greek
term p3ptidia translated as small digestibles or digested
(Shahidi & Zhong, 2008). Biological activity of peptides were rst
reported by Mellander in 1950. In the last two decades there has
been a strong interest in bioactive peptide identication and
characterization. Researchers continue to investigate new sources, extraction methods and evaluate health benets of bioactive
peptides. Bioactive peptides are specic protein fragments that
can alter body functions or conditions and may ultimately inuence health positively (Korhonen & Pihlanto, 2006). Such
peptides are inactive within the sequence of the parent protein
and can be released by several hydrolysis techniques. Bioactive
peptides have potential to help reduce the worldwide epidemic
of chronic diseases affecting 58 million people annually (Sun,
* Corresponding author.
E-mail address: Brijesh.tiwari@teagasc.ie (B.K. Tiwari).
http://dx.doi.org/10.1016/j.tifs.2015.07.012
0924-2244/ 2015 Elsevier Ltd. All rights reserved.
2013). Currently the market for functional proteins and peptides is valued at $75 billion/year (Sun, 2013). Peptide based
drugs sales are growing at a rapid pace with annual sales of $20
billion corresponding to 2% of the global drug market (Sun,
2013). Bioactive peptides from food proteins offer major potential for incorporation into functional foods and nutraceuticals.
Major drivers for bioactive peptides drugs include their role as
hormones, neurotransmitters and neuromodulaters, and low
toxicity or non-toxicity of metabolic cleavage products. Challenges associated with the introduction of new peptide products
include proteolytic degradation, fast clearance in the body, low
solubility in water, immunogenicity (Bellmann-Sickert & BeckSickinger, 2010) and regulatory hurdles. Inactive or latent form
of bioactive peptides in parent protein can be activated by proteolysis (Ryan, Ross, Bolton, Fitzgerald, & Stanton, 2011). Bioactive peptides may be produced by several methods: the most
commonly employed are based on enzymatic hydrolysis by
digestive enzymes derived from micro-organisms and fermentation of food proteins (Kim & Wijesekara, 2013). However other
methods based on chemical hydrolysis or sub-critical water hydrolysis have also been widely researched in recent decades
(Rogalinski, Herrmann, & Brunner, 2005).
S.U. Kadam et al. / Trends in Food Science & Technology 46 (2015) 60e67
61
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S.U. Kadam et al. / Trends in Food Science & Technology 46 (2015) 60e67
classes. Peptides can also be classied on the basis of their bioactivity. The applications of peptides have evolved from being a
simple source of nitrogen and energy towards applications in human health (Fig 3). Peptides possess diverse biological activities
such as opiate, antithrombotic, anti-hypertensive, immunomodulating, antilipemic, osteoprotective, antioxidative, antimicrobial,
Fig. 3. Evolution of peptides and applications. Modied from (Sharma, Singh, & Rana, 2011).
S.U. Kadam et al. / Trends in Food Science & Technology 46 (2015) 60e67
63
Table 1
Ultrasound mechanisms for different applications.
Application
Ultrasound mechanism
References
Protein digestion
Ultrasonic bubble cavitation causes rapid changes of temperature, pressure, shear forces, jets, and shock waves
which results in mixing of reaction components instantly and proteolysis
Implosion of cavitational bubbles, formation of microjets, micro-turbulence, high-velocity inter-particle collisions
and perturbation in micro-porous particles results in enhanced extraction yield and accelerated chemical reactions
Increase in the surface hydrophobicity and loosening of protein for easy release of peptides
pez-Ferrer et al.,
(Lo
2008)
(Shirsath et al., 2012)
Isolation/Extraction of
protein
Production of ACE
inhibitory peptide
Encapsulation of
bioactive peptides
Sonophoresis
64
S.U. Kadam et al. / Trends in Food Science & Technology 46 (2015) 60e67
S.U. Kadam et al. / Trends in Food Science & Technology 46 (2015) 60e67
combinations thereof (Marczak et al., 2003). ACE inhibitory peptides, can be divided into three groups, namely substrate, inhibitor
and prodrug type due to presence of ACE substrates in enzymatic
food protein digests (Iroyukifujita, Eiichiyokoyama, & Yoshikawa,
2000). An overview of ACE-inhibitory peptides is outlined in
Fig. 5. Jia et al. (2010) used ultrasound pre-treatment to enhance the
breakdown of defatted wheat germ protein to yield ACE inhibitor
peptides. On average the breakdown rate constant increased by
22.2% when treated with ultrasound at 22 kHz frequency and 40 W
for 210 min. Ultrasound treatment also elevated ACE inhibitor activity levels. Zhou et al. (2013) reported that an ultrasound probe
system can deliver better ACE inhibitor activity compared to an
ultrasonic bath system.
Ultrasonic-assisted hydrolysis at a power level of 150 W for
25 min treatment can be used to improve the antioxidant activities
of peanut protein (Yu et al., 2012). Ultrasound at an intensity of
0.707 W cm2 and frequency of 25 kHz was employed to produce
wheat gluten hydrolysates with enhanced iron-chelating, antioxidant and ABTS radical scavenging activities (Zhu, Su, Guo, Peng, &
Zhou, 2011). The same authors reported that molecular weight
distribution was strongly inuenced by ultrasonic frequency, being
lower in size when a lower frequency was applied. Peptides smaller
than 1 kDa possess the best antioxidant properties (Moure,
, 2006); thus the desirable production of low
Domnguez, & Parajo
molecular weight peptides can be facilitated using ultrasound.
3.4. Drug delivery
In general peptide based drugs are physically and chemically
unstable, have short in vivo half-lifes and have low oral bioavailability. Encapsulation of peptide based drugs with biodegradable
polymers can improve stability and bioavailability.
There are number of encapsulation techniques used for protein
65
drug delivery including ultrasonic atomization, electrospray, micro-uidic, pore-closing, thermo-reversible-gel, and microfabrication (Ye, Kim, & Park, 2010). Ultrasonic atomization is a
simple and continuous technique, and aseptic in nature (Ye et al.,
2010). Ultrasonic atomization is the process of breaking up of
thin lm of liquid into ne droplets when allowed to ow on a
vibrating surface (frequency >20 kHz). Capillary wave and cavitation are the two main hypotheses used to explain the mechanism of
liquid disintegration during ultrasonic atomization. The cavitation
hypothesis is generally applied to high frequency and high-energy
intensity systems. During the implosive collapse of cavities formed
during sonication, high intensity hydraulic shocks are generated.
These hydraulic shocks initiate the disintegration of the liquid lm
and cause direct ejection of the droplets (Avvaru, Patil, Gogate, &
Pandit, 2006). Various ultrasonic atomizer systems have been
developed over the years to produce protein microspheres (Freitas,
Rudolf, Merkle, & Gander, 2005; Yeo, Chen, Basaran, & Park, 2004).
Liposomes, composed of a lipid bilayer, are generally used as protein delivery carriers due to their robust and convenient nature to
deliver large protein molecules (Lu et al., 2010). However,
microbubble-enhanced ultrasound can be more efcient than liposomes for controlled release of proteins and peptides (Kinoshita
& Hynynen, 2005).
Sonophoresis is the use of ultrasound technology at a frequency
of 20 kHz-16 MHz and power intensity of up to 14 W cm2 to increase the penetration of molecules into and across the skin (Park,
Park, Seo, & Lee, 2014). Permeability of skin is enhanced by various
cavitational and non-cavitational mechanisms of ultrasound. In
high-frequency ultrasound (0.7e16 MHz) cavitation occurs within
the skin i.e. within cavities near the corneocytes of the stratum
corneum. However, low-frequency ultrasound (<100 kHz) uses
transient cavitation inside and outside the skin. Convection, thermal effects, mechanical or radiation pressure effects, lipid
MECHANISM
SOURCES
Chicken eggs,
Wheat, Corn,
Garlic, Milk,
Fish
KininKalicrein
System
ACE-Inhibitory
Pepdes
Substrate
Inhibitor
Prodrug
Producon
Tradional
- Protein hydrolysis by
using enzymes such as
pepsin, pancrean,
trypsin, chymotrypsin,
thermolysin, sublisin
and combinaons of
pepsin with other
enzymes
Ultrasound
- Improved dispersion and
less agglomeraon of
enzymes.
- Higher acvity
- Increased producon rates
- Probe ultrasound beer
than bath ultrasound
system
TYPES
ReninAngiotensin
System
66
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S.U. Kadam et al. / Trends in Food Science & Technology 46 (2015) 60e67
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