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Journals / Türk Biyokimya Dergisi / 2018 / Cilt: 43 - Sayı: 6

Purification and biochemical characterization of a β-cyanoalanine synthase expressed in germinating seeds of Sorghum bicolor (L.) moench

Sorghum bicolor (L.) Moench’in Çimlenen Tohumlarında İfade Edilen Bir β-Siyanoalanin Sentazın Saflaştırılması ve Biyokimyasal Karakterizasyonu

Ruth Ololade AMIOLA
Adedeji Nelson ADEMAKIMWA
Zainab Adenike AYINLA
Esther Nkechi EZIMA
Femi Kayode AGBOOLA
Pages
638–650
Publication Date
2022-11-05
DOI
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AI Summary & Key Takeaways

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⚡ TL;DR(Too Long; Didn't Read)

Inhibitorssuch as HgCl2, EDTA, glycine and iodoacetamide reducedenzyme activity.Conclusion: The biochemical properties of β-cyanoalaninesynthase in germinating sorghum seeds highlights itsroles in maintaining cyanide homeostasis.

📌 3 Key Takeaways

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Objective & Hypothesis:Background: β-Cyanoalanine synthase plays essentialroles in germinating seeds, such as in cyanide homeostasis.Methods: β-Cyanoalanine synthase was isolated fromsorghum seeds, purified using chromatographic techniquesand its biochemical and catalytic properties weredetermined.Results: The purified enzyme had a yield of 61.74% andspecific activity of 577.50 nmol H2S/min/mg of protein.The apparent and subunit molecular weight for purifiedβ-cyanoalanine synthase were 58.26 ± 2.41 kDaand 63.4 kDa, respectively.
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Key Findings & Data:The kinetic parameterswith sodium cyanide as substrate were 0.67 ± 0.08 mM,17.60 ± 0.50 nmol H2S/mL/min, 2.97 × 10−1 s−1 and4.43 × 102 M−1 s−1 for KM, Vmax, kcat and kcat/KM, respectively.With L-cysteine as substrate, the kinetic parameterswere 2.64 ± 0.37 mM, 63.41 ± 4.04 nmol H2S/mL/min,10.71 × 10−1 s−1 and 4.06 × 102 M−1 s−1 for KM, Vmax, kcat andkcat/KM, respectively.
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Conclusion & Impact:Inhibitorssuch as HgCl2, EDTA, glycine and iodoacetamide reducedenzyme activity.Conclusion: The biochemical properties of β-cyanoalaninesynthase in germinating sorghum seeds highlights itsroles in maintaining cyanide homeostasis.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:Domain Parameters

Abstract

Background: β-Cyanoalanine synthase plays essentialroles in germinating seeds, such as in cyanide homeostasis.Methods: β-Cyanoalanine synthase was isolated fromsorghum seeds, purified using chromatographic techniquesand its biochemical and catalytic properties weredetermined.Results: The purified enzyme had a yield of 61.74% andspecific activity of 577.50 nmol H2S/min/mg of protein.The apparent and subunit molecular weight for purifiedβ-cyanoalanine synthase were 58.26 ± 2.41 kDaand 63.4 kDa, respectively. The kinetic parameterswith sodium cyanide as substrate were 0.67 ± 0.08 mM,17.60 ± 0.50 nmol H2S/mL/min, 2.97 × 10−1 s−1 and4.43 × 102 M−1 s−1 for KM, Vmax, kcat and kcat/KM, respectively.With L-cysteine as substrate, the kinetic parameterswere 2.64 ± 0.37 mM, 63.41 ± 4.04 nmol H2S/mL/min,10.71 × 10−1 s−1 and 4.06 × 102 M−1 s−1 for KM, Vmax, kcat andkcat/KM, respectively. The optimum temperature and pHfor activity were 35°C and 8.5, respectively. The enzymeretained more than half of its activity at 40°C. Inhibitorssuch as HgCl2, EDTA, glycine and iodoacetamide reducedenzyme activity.Conclusion: The biochemical properties of β-cyanoalaninesynthase in germinating sorghum seeds highlights itsroles in maintaining cyanide homeostasis.

Özet

Giriş: β-siyanoalanin sentaz, çimlenen tohumlarda, siyanür homeostazı gibi önemli roller oynamaktadır. Yöntemler: β-siyanoalanin sentaz, sorgum tohumlarından izole edildi, kromatografik teknikler kullanılarak saflaştırıldı ve biyokimyasal ve katalitik özellikleri belirlendi. Bulgular: Saflaştırılmış enzim,% 61.74’lük bir verim ve 577.50 nmol H2S/dak/mg protein spesifik aktivitesine sahipti. Saflaştırılmış β-siyanoalanin sentaz için görünür ve alt birim molekül ağırlığı sırasıyla 58.26 ± 2.41 kDa ve 63.4 kDa idi. Substrat olarak sodyum siyanür ile kinetik parametreler, KM, Vmax, kcat ve kcat/KM için sırasıyla 0.67 ± 0.08 mM, 17.60 ± 0.50 nmol H2S/mL/dak, 2.97 × 10−1 s−1 ve 4.43 × 102 M−1 s−1 olarak belirlendir. Substrat olarak L-sistein ile kinetik parametreler KM, Vmax, kcat ve kcat/KM için sırasıyla 2.64 ± 0.37 mM, 63.41 ± 4.04 nmol H2S/mL/ dak, 10.71 × 10−1 s−1 ve 4.06 × 102 M−1 s−1 idi. Aktivite için optimum sıcaklık 35°C ve optimum pH 8.5 idi. Enzim, aktivitesinin yarısından fazlasını 40°C’de korudu. HgCl2, EDTA, glisin ve iyodoasetamid gibi inhibitörler enzim aktivitesini düşürdü. Sonuç: Çimlenmekte olan sorgum tohumlarındaki β-siyanoalanin sentazın biyokimyasal özellikleri, siyanür homeostazının korunmasındaki rolünü vurgulamaktadır.

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Cite This Article

AMIOLA, R. O., ADEMAKIMWA, A. N., AYINLA, Z. A., EZIMA, E. N., AGBOOLA, F. K. (2022). Purification and biochemical characterization of a β-cyanoalanine synthase expressed in germinating seeds of Sorghum bicolor (L.) moench. Türk Biyokimya Dergisi, Cilt: 43 - Sayı: 6, 638–650. https://ajindex.com/articles/620951

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References (18)

  1. Conn EE. Cyanogenic glycosides. New York: Academic Press, 1981:479–500
  2. Møller BL. Functional diversifications of cyanogenic glucosides. Curr Opin Plant Biol 2010;13:338–47
  3. Lechtenberg M. Cyanogenesis in higher plants and animals. Chichester: John Wiley and Sons, 2011
  4. Selmar D, Lieberei R, Biehl B. Mobilization and utilization of cyanogenic glycosides. The linustatin pathway. Plant Physiol 1988;86:711–6
  5. Vetter J. Plant cyanogenic glycosides. Toxicon 1988;38:11–36
  6. Kende H. Enzymes of ethylene biosynthesis. Plant Physiol 1989;91:1–4
  7. Seigler DS. Cyanide and cyanogenic glycosides. In: Herbivores: their interactions with secondary plant metabolites. San Diego, CA: Academic Press, 1991:35–77
  8. Kadow D, Voß K, Selmar D, Lieberei R. The cyanogenic syndrome in rubber tree Hevea brasiliensis: tissue-damage-dependent activation of linamarase and hydroxynitrile lyase accelerates hydrogen cyanide release. Ann Bot 2012;109:1253–62
  9. Widmer TL, Abawi GS. Relationship between levels of cyanide in sudangrass hybrids incorporated into soil and suppression of Meloidogyne hapla. J Nematol 2002;34:16–22
  10. Peiser GD, Tsu-Tsuen W, Hoffman NE, Shang Y, Liu HW, Walsh CT. Formation of cyanide from carbon 1 of 1-aminocyclopropane- 1-carboxylic acid during its conversion to ethylene. Proc Natl Acad Sci USA 1984;81:3059–63
  11. Manning K. Ethylene production and β-cyanoalanine synthase activity in carnation flowers. Planta 1986;168:61–6
  12. Morgan PW, Drew MC. Ethylene and plant responses to stress. Physiol Plant 1997;100:620–30
  13. Seo S, Mitsuhara I, Feng J, Iwai T, Hasegawa M, Ohashi Y. Cyanide, a co-product of plant hormone ethylene biosynthesis, contributes to the resistance of rice to blast fungus. Plant Physiol 2011;155:502–14
  14. Woodrow IE, Slocum DJ, Gleadow RM. Influence of water stress on cyanogenic capacity in Eucalyptus cladocalyx. Funct Plant Biol 2002;29:103–10
  15. Liang WS. Drought stress increases both cyanogenesis and β-cyanoalanine synthase activity in tobacco. Plant Science 2003;165:1109–15
  16. Donato DB, Nichols O, Possingham H, Moore M, Ricci PF, Noller BN. A critical review of the effects of gold cyanide-bearing tailings solutions on wildlife. Environ Int 2007;33:974–84
  17. Samiotakis M, Ebbs SD. Possible evidence for transport of an iron cyanide complex by plants. Environ Pollut 2004;127:169–73
  18. Gupta N, Balomajumder C, Agarwal VK. Enzymatic mechanism and biochemistry for cyanide degradation: a review. J Hazard Mater