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Protein Extraction and Identification by Gel Electrophoresis and Mass Spectrometry from Edible bird’s Nest Samples

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Abstract

Edible bird’s nest (EBN) is a delicacy rich in protein and carbohydrate from the salivary secretion of swiftlets. There are limited studies on the protein profile of EBN, mainly due to its complexity in chemical composition and diversity of species, as well as the capacity of analytical techniques. The protein extraction methods, namely, ultrasonic and detergent (Triton X-100 and SDS)-assisted methods, as well as Tris–HCl buffer solubilization were used to compare the protein profiles of EBNs harvested from two locations (Batu Pahat and Kota Tinggi) in Malaysia. Ultrasonic assisted extraction produced the highest protein content because EBN contains mostly water-soluble protein. The electrophoretic gels revealed that EBNs from Batu Pahat (17–150 kDa) exhibited more protein bands than those samples from Kota Tinggi (25–154 kDa). The difference could be explained by the variance in the geographical origin of EBNs. Additional protein bands (25, 27, and 92 kDa) were detected for the detergent-assisted methods. These could be poorly water-soluble membrane proteins that were released after cell lysis by detergents. The mass spectra revealed that acidic mammalian chitinase precursor is the most abundant protein. The other proteins include trypsinogen, inter-alpha-trypsin inhibitor heavy chain H2 precursor, tumor necrosis factor receptor superfamily member 5 precursor, and phospholipase A2-like. There are also collagen and co-enzyme Q-binding proteins which are important for skin complexion in line with the traditional belief of Chinese community. The protein extraction methods could produce good quality of proteins for affirmative confirmation using gel electrophoresis and mass spectrometric identification.

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References

  • Bahuguna R, Purohit MC, Nautiyal BP, Prakash V, Nautiyal MC, Purohit AN (2003) Polypeptide patterns and isoenzymes in different populations of Aconitum heterophyllum wall ex Royle, from Garhwal Himalaya. Physiol Mol Biol Plants 9:95–100

    Google Scholar 

  • Berezovski MV, Mak TW, Krylov SN (2007) Cell lysis inside the capillary facilitated by transverse diffusion of laminar flow profiles (TDLFP). Anal Bioanal Chem 387:91–96

    Article  CAS  Google Scholar 

  • Bollag DM, Rozycki MD, Edelstein SJ (1996) Protein methods: second edition. Wiley. Publication, New York

  • Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  • Brown RB, Audet J (2008) Current techniques for single-cell lysis. J Roy Soc Interface 5:S131–S138

    Article  CAS  Google Scholar 

  • Chan KM, Zhang Z (2012) Leucine-rich repeat and WD repeat-containing protein 1 is recruited to pericentric heterochromatin by trimethylated lysine 9 of histone H3 and maintains heterochromatin silencing. J Biol Chem 287:15024–15033

    Article  CAS  Google Scholar 

  • Chua KH, Lee TH, Kamini N, Nor HMY, Lee CT, Eddie TTT, Ramlan AA (2013) Edible Bird’s Nest as a chondro-protective agent for human chondrocytes isolated from osteoarthritic knee: in vitro study. BMC Complement Altern Med 13:19

    Article  Google Scholar 

  • Chua LS, Nurulaini AR, Bustanur R, Lee CT (2012) Plant proteins, minerals and trace elements of Eurycoma longifilia (Tongkat ali). Nat Prod Res 27(4–5):314–318

    Google Scholar 

  • Contreras L, Ritter A, Dennett G, Boehmwald F, Guitton N, Pineau C, Moenne A, Potin P, Correa JA (2008) Two dimensional gel electrophoresis analysis of brown algal protein extracts. J Phycol 44:1315–1321

    Article  CAS  Google Scholar 

  • Dennis EA, Cao J, Hsu YH, Magrioti V, Kokotos G (2011) Phospholipase A2 enzymes: physical structure, biological function, disease implication, chemical inhibition, and therapeutic intervention. Chem Rev 111:6130–6185

    Article  CAS  Google Scholar 

  • Fraser JR, Laurent TC, Laurent UB (1997) Hyaluronan: its nature, distribution, functions and turnover. J Intern Med 242(1):27–33

    Article  CAS  Google Scholar 

  • Friedenauer S, Berlet HH (1989) Sensitivity and variability of the Bradford protein assay in the presence of detergents. Anal Biochem 78:263–268

    Article  Google Scholar 

  • Guo CT, Takahashi T, Bukawa W, Takahashi N, Yagi H, Kato K, Miyamoto D, Suzuki D, Suzuki Y (2006) Edible bird’s nest extract inhibits influenza virus infection. Antivir Res 70:140–146

    Article  CAS  Google Scholar 

  • Heringdorf DM, Jakobs KH (2007) Lysophospholipid receptors: signalling, pharmacology and regulation by lysophospholipid metabolism. Biochim Biophys Acta 1768:923–940

    Article  Google Scholar 

  • Herrera-Estrella A, Chet I (1999) Chintinases in biological control. In: Jolles P, Muzzarelli RAA (eds) Chitin and chitinase. Birkhauser, Basel, pp. 171–184

    Chapter  Google Scholar 

  • Kang N, Hails CJ, Sigurdsson JB (1991) Nest construction and egg-laying in edible-nest swiflets Aerodramus spp. and the implications for harvesting. Ibis 133(2):170–177

    Article  Google Scholar 

  • Kathan RH, Weeks DI (1969) Structure studies of collocalia mucoid: I. Carbohydrate and amino acid composition. Arch Biochem Biophys 134:572–576

    Article  CAS  Google Scholar 

  • Kechrid Z, Bouzerna N (2004) Effect of zinc deficiency on zinc and carbohydrate metabolism in genetically diabetic C57BL/Ksj Db+/Db+) and non-diabetic original strain (C57BL/Ksj) mice. Turk J Med Sci 34:367–373

    CAS  Google Scholar 

  • Kong YC, Keung WM, Yip TT, Ko KM, Tsao SW, Ng MH (1987) Evidence that epidermal growth factor is present in swiftlet’s (Collocalia) nest. Comp Biochem Physiol Biochem Mol Biol 87:221–226

    Article  CAS  Google Scholar 

  • Koon LC, Cranbrook E (2002) Swiftlets of Borneo—builders of edible nests. Natural History Publication (Borneo) SDN. BHD., Sabah, pp. 1–171

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  • Liu X, Lai X, Zhang S, Huang X, Lan Q, Li Y, Li BC, Zhang Q, Hong D, Yang G (2012) Proteomic profile of edible bird’s nest. J Agric Food Chem 60:12477–12481

    Article  CAS  Google Scholar 

  • Ma F, Liu D (2012) Sketch of the edible bird’s nest and its important bioactivities. Food Res Int 48:559–567

    Article  CAS  Google Scholar 

  • Marcone MF (2005) Characterization of the edible bird’s nest the “Caviar of the East”. Food Res Int 38:1125–1134

    Article  CAS  Google Scholar 

  • Marni S, Marzura MR, Norzela AM, Khairunnisak M, Bing CH, Eddy AA (2014) Preliminary study on free sialic acid content of edible bird nest from Johor and Kelantan. Malaysian J Vet Res 5:9–14

    Google Scholar 

  • Matsukawa N, Matsumoto M, Bukawa W, Chiji H, Nakayama K, Hara H, Tsukahara T (2011) Improvement of bone strength and dermal thickness due to dietary edible bird’s nest extract in ovariectomized rats. Biosci Biotechnol Biochem 75:590–592

    Article  CAS  Google Scholar 

  • Murphy RM, Lamb GD (2013) Important considerations for protein analyses using antibody based techniques: down-sizing Western blotting up-sizes outcomes. J Physiol 591(Pt 23):5823–5831

    Article  CAS  Google Scholar 

  • Nakagawa H, Hama Y, Sumi T, Li SC, Maskos K, Kalayanamitra K, Mizumoto S, Sugahara K, Li YT (2007) Occurrence of a non-sulfated chondroitin proteoglycan in the dried saliva of Collocalia swiftlets (edible bird’s-nest). Glycobiology 17:157–164

    Article  CAS  Google Scholar 

  • Qiu R, Yang Y, Zhao H, Li J, Xin Q, Shan S, Liu Y, Dang J, Yu X, Gong Y, Liu Q (2013) Signal transducer and activator of transcription 6 directly regulates human ORMDL3 expression. FEBS J 280:2014–2026

    Article  CAS  Google Scholar 

  • Rajapakse S, Ogiwara K, Takahashi T (2014) Characterization and expression of trypsinogen and trypsin in medaka testis. Zool Sci 31(12):840–848

    Article  Google Scholar 

  • Roh KB, Lee J, Kim YS, Park J, Kim JH, Lee J, Park D (2012) Mechanisms of edible bird’s nest extract-induced proliferation of human adipose-derived stem cells. Evid Based Complement Alt Med 2012:Article ID 797520, 11 pages.

  • Sazili AQ, Noor Azihan MZ, Shuhaimi M, Hilmi M, Panandam JM (2010) The effectiveness of soluble protein extractability under the effect of pH, molarity, type of buffers of three different major skeletal muscles in cattle. Asian J Biol Sci 1–5

  • Shevchenko A, Wilm M, Vorm O, Mann M (1996) Mass spectrometric sequencing of proteins from silver stained polyacrylamide gels. Anal Chem 68:850–858

    Article  CAS  Google Scholar 

  • Sims RW (1961) The identification of Malaysian species of swiftlets. Ibis 103a(2):205–210

    Article  Google Scholar 

  • Utomo B, Rosyidi D, Radiati LE, Puspaningsih NNT, Proborini WD (2014) Protein characterization of extracted water from three kinds of edible bird nest using SDS-PAGE CBB staining and SDS-PAGE glycoprotein staining and LC-MS/MS analyses. IOSR J Agr Vet Sci 7:33–38

    Article  Google Scholar 

  • Vimala B, Hussain H, Wan Nazaimoon WM (2011) Effects of edible bird’s nest on tumour necrosis factor-alpha secretion, nitric oxide production and cell viability of lipopolysaccharide-stimulated RAW 264.7 macrophages. Food Agric Immunol 23:303–314

    Article  Google Scholar 

  • Yan H, Lamm ME, Bjorling E, Huang YT (2002) Multiple functions of immunoglobulin A in mucosal defense against viruses: an in vitro measles virus model. J Virol 76(21):10972–10979

    Article  CAS  Google Scholar 

  • Yew MY, Koh RY, Chye SM, Othman I, Ng KY (2014) Edible bird’s nest ameliorates oxidative stress-induced apoptosis in SH-SY5Y human neuroblastoma cells. BMC Complement Altern Med 2014(14):391

    Article  Google Scholar 

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Authors and Affiliations

  1. Metabolites Profiling Laboratory, Institute of Bioproduct Development, Universiti Teknologi Malaysia, 81310 UTM, Johor Bahru, Johor, Malaysia

    Siti Najihah Zukefli & Lee Suan Chua

  2. Department of Bioprocess and Polymer Engineering, Faculty of Chemical and Energy Engineering, Universiti Teknologi Malaysia, 81310 UTM, Johor Bahru, Johor, Malaysia

    Siti Najihah Zukefli & Lee Suan Chua

  3. Department of Biotechnology and Medical Engineering, Faculty of Biosciences and Medical Engineering, Universiti Teknologi Malaysia, 81310 UTM, Johor Bahru, Johor, Malaysia

    Zaidah Rahmat

Authors
  1. Siti Najihah Zukefli
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  2. Lee Suan Chua
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  3. Zaidah Rahmat
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Correspondence to Lee Suan Chua.

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Funding

This study was funded by Longevity Wellness Industries Sdn Bhd. (4C046).

Conflict of Interest

S.N. Zukefli declares that she has no conflict of interest. L.S. Chua declares that she has no conflict of interest. Z. Rahmat declares that she has no conflict of interest.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Zukefli, S.N., Chua, L.S. & Rahmat, Z. Protein Extraction and Identification by Gel Electrophoresis and Mass Spectrometry from Edible bird’s Nest Samples. Food Anal. Methods 10, 387–398 (2017). https://doi.org/10.1007/s12161-016-0590-7

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  • DOI: https://doi.org/10.1007/s12161-016-0590-7

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