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Bioconversion of bread waste by marine psychrotolerant Glutamicibacter soli strain AM6 to a value-added product: cold-adapted, salt-tolerant, and detergent-stable α-amylase (CA-AM21)

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Abstract

The present work aims to exploit the zero-cost inducer bread waste (BW) for the production of cold-adapted α-amylase (CA-AM21) by the hyperproducer marine psychrotolerant Glutamicibacter soli strain AM6 EMCCN 3074. Incubation temperature (20 °C), incubation time, BW, (NH4)2SO4, and modified seawater-based minimal medium were inferred to be significant factors influencing CAM-21 production in OVAT and Plackett-Burman investigations. BW (3.8%w/v), (NH4)2SO4 (0.098%w/v), and incubation time (2.329 days) prompted CA-AM21 (56.85 U/mL) with 7.58-fold enhancement, Box-Behnken design, and ridge steepest ascent path inferences. A two-step strategy was used to purify CA-AM21: fractional (NH4)2SO4 precipitation (60-80%) and a starch affinity chromatography. Specific activity, yield, and fold purification of purified CA-AM21 were 884.5 U/mg, 12.45%, and 285.3, respectively. CA-AM21 exhibited an approximate molecular mass of 14 kDa, deduced from SDS-PAGE analyses. The pH and temperature optimums for CA-AM21 were 7.0 and 40 °C, respectively. At 5–25 °C, full activity was maintained after 2 h. At pH 6.0–8.0, almost full activity (90–100%) was retained after 15 h. CA-AM21 demonstrated full stability after 30 min at 1.0–5.0 M NaCl, with 200% increased activity at 1.0–4.0 M NaCl. Full stability of CA-AM21 in the presence of non-ionic detergents, such as Tween 20, Tween 80, and Triton X-100, with 100% activity sustained after 30 min was retained. On soluble starch, purified CA-AM21 exhibited km and Vmax of 0.039 mg.mL−1 and 115.71 μmol glucose. min−1. mg−1, respectively. The purified CA-AM21 demonstrated remarkable significant washing efficiency with commercial laundry detergents thus implying its applicability for detergent formulations.

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References

  1. Kuddus M (2014) Cold-active microbial enzymes. Biochem Physiol Open Access 04. https://doi.org/10.4172/2168-9652.1000e132

  2. Nandanwar S, Borkar S, Lee J, Kim HJ (2020) Taking advantage of promiscuity of cold-active enzymes Appl Sci 10:8128. https://doi.org/10.3390/app10228128

    Article  Google Scholar 

  3. van der Maarel MJEC, van der Veen B, Uitdehaag JCM, Leemhuis H, Dijkhuizen L (2002) Properties and applications of starch-converting enzymes of the alpha-amylase family. J Biotechnol 94:137–155. https://doi.org/10.1016/s0168-1656(01)00407-2

    Article  Google Scholar 

  4. Maharana A, Singh S (2018) Cold active amylases producing psychrotolerants isolated from Nella Lake, Antarctica Biosci Biotechnol Res Asia 15. 10.13005/bbra/2603

  5. Kuddus M, Saima R, Ahmad I (2012) Cold-active extracellular α-amylase production from novel bacteria Microbacterium foliorum GA2 and Bacillus cereus GA6 isolated from Gangotri glacier, Western Himalaya. J Genet Eng Biotechnol 10:151–159. https://doi.org/10.1016/j.jgeb.2012.03.002

    Article  Google Scholar 

  6. Morita RY (1975) Psychrophilic bacteria. Bacteriol Rev 39:144–167. https://doi.org/10.1128/br.39.2.144-167.1975

    Article  Google Scholar 

  7. Kim S, Park H, Choi J (2017) Cloning and characterization of cold-adapted α-amylase from Antarctic Arthrobacter agilis. Appl. Biochem Biotechnol 181:1048–1059. https://doi.org/10.1007/s12010-016-2267-5

    Article  Google Scholar 

  8. Roohi KM (2014) Bio-statistical approach for optimization of cold-active α-amylase production by novel psychrotolerant M. foliorum GA2 in solid state fermentation. Biocatal Agri Biotechnol 3:175–181. https://doi.org/10.1016/j.bcab.2013.09.007

    Article  Google Scholar 

  9. Roohi R, Kuddus M, Saima S (2013) Cold-active detergent-stable extracellular α-amylase from Bacillus cereus GA6: biochemical characteristics and its perspectives in laundry detergent formulation. J Biochem Technol 4:636–644

    Google Scholar 

  10. Hamid DB (2015) Cold-active α-amylase from psychrophilic and psychrotolerant yeast. J Glob Biosci 4:2670–2677

    Google Scholar 

  11. Fenice M, Selbmann L, Zucconi L, Onofri S (1997) Production of extracellular enzymes by Antarctic fungal strains. Polar Biol 17:275–280. https://doi.org/10.1007/s003000050132

    Article  Google Scholar 

  12. Melikoglo M (2008) Production of sustainable alternatives to petrochemicals and fuels using waste bread as a raw material. The University of Manchester

    Google Scholar 

  13. Leung C, Cheung A, Zhang A, Lam K, Lin C (2012) Utilisation of waste bread for fermentative succinic acid production. Biochem Eng J 65:10–15. https://doi.org/10.1016/j.bej.2012.03.010

    Article  Google Scholar 

  14. Han W, Hu Y, Li S, Huang J, Nie Q, Zhao H, Tang J (2017) Simultaneous dark fermentative hydrogen and ethanol production from waste bread in a mixed packed tank reactor. J Clean Prod 141:608–611. https://doi.org/10.1016/j.jclepro.2016.09.143

    Article  Google Scholar 

  15. Cerda A, El-Bakry M, Gea T, Sánchez A (2016) Long term enhanced solid-state fermentation: inoculation strategies for amylase production from soy and bread wastes by Thermomyces sp. in a sequential batch operation. J Environ Chem Eng:4. https://doi.org/10.1016/j.jece.2016.04.022

  16. Gómez-Villegas P, Vigara J, Romero L, Gotor C, Raposo S, Gonçalves B, Léon R (2021) Biochemical characterization of the amylase activity from the new haloarchaeal strain Haloarcula sp. HS isolated in the Odiel marshlands. Biology 10: 337.

  17. Männistö MK, Puhakka JA (2002) Psychrotolerant and microaerophilic bacteria in boreal groundwater. FEMS microbiology ecology 41(1):9–16

    Article  Google Scholar 

  18. Eden PA, Schmidt TM, Blakemore RP, Pace NR (1991) Phylogenetic analysis of Aquaspirillum magnetotacticum using polymerase chain reaction amplified 16S rRNA-specific DNA. Int J Syst Bacteriol 41:324–325

    Article  Google Scholar 

  19. Embaby AM, Heshmat Y, Hussein A, Marey HS (2014) A sequential statistical approach towards an optimized production of a broad spectrum bacteriocin substance from a soil bacterium Bacillus sp. YASI strain. Sci World J 2014:396304

    Google Scholar 

  20. Asgher M, Asad MJ, Rahman SU, Legge R. L (2007) A thermostable α-amylase from a moderately thermophilic Bacillus subtilis strain for starch processing. J Food Eng 79: 950–955. https://doi.org/10.1016/j.jfoodeng.2005.12.053.

  21. Nguyen T (2018) Preparation of artificial sea water (ASW) for culturing marine bacteria. 10.13140/RG.2.2.20641.71528.

  22. Okolo BN, Ezeogu LI, Mba CN (1995) Production of raw starch digesting amylase by Aspergillus niger grown on native starch sources. J. Sci. Food Agric 69:109–115

    Article  Google Scholar 

  23. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428. https://doi.org/10.1021/ac60147a030

    Article  Google Scholar 

  24. Plackett RL, Burman JP (1946) The design of optimum multifactorial experiments. Biometrika 33:305–325

    Article  MathSciNet  MATH  Google Scholar 

  25. Box GEP, Behnken DW (1960) Some new three level designs for the study of quantitative variables. Technometrics 2:455–475

    Article  MathSciNet  Google Scholar 

  26. Rao MD, Ratnam BVV, VenkataRamesh D, Ayyanna C (2005) Rapid method for the affinity purification of thermostable α-amylase from Bacillus licheniformis. World J Microbiol Biotechnol 21:371–375. https://doi.org/10.1007/s11274-004-3908-3

    Article  Google Scholar 

  27. Bradford MM (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  Google Scholar 

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

    Article  Google Scholar 

  29. Liu G, Wu S, Jin W, Sun C (2016) Amy63, a novel type of marine bacterial multifunctional enzyme possessing amylase, agarase and carrageenase activities. Sci Rep 6(1):1-. https://doi.org/10.1038/srep18726.

  30. Shukla RJ, Singh SP (2015) Characteristics and thermodynamics of α-amylase from thermophilic actinobacterium, Laceyella sacchari TSI-2. Process Biochem 50:2128–2136. https://doi.org/10.1016/j.procbio.2015.10.013

    Article  Google Scholar 

  31. Yemm EW, Willis A (1954) The estimation of carbohydrates in plant extracts by anthrone. Biochem J 57(3):508–514

    Article  Google Scholar 

  32. Myers RH (1976) Response surface methodology. Edwards Brothers, Ann Arbor, MI

    Google Scholar 

  33. Draper NR (1963) “Ridge analysis” of response surfaces. Technometrics 5:469–479

    MATH  Google Scholar 

  34. Matrawy AA, Khalil AI, Marey HS, Embaby AM (2020) Biovalorization of the raw agro-industrial waste rice husk through directed production of xylanase by Thermomyces lanuginosus strain A3-1 DSM 105773: a statistical sequential model. Biomass Convers Biorefinery. https://doi.org/10.1007/s13399-020-00824-9

  35. Embaby AM, Hussein MN, Hussein A (2018) Monascus orange and red pigments production by Monascus purpureus ATCC16436 through co-solid state fermentation of corn cob and glycerol: an eco-friendly environmental low cost approach. PLoS One 13:e0207755

    Article  Google Scholar 

  36. Anan A, Ghanem KM, Embaby AM, Hussein A, El-Naggar MY (2018) Statistically optimized ceftriaxone sodium biotransformation through Achromobacter xylosoxidans strain Cef6: an unusual insight for bioremediation. J Basic Microbiol 58:120–130. https://doi.org/10.1002/jobm.201700497

    Article  Google Scholar 

  37. Embaby AM, Melika RR, Hussein A, El-Kamel AH, S Marey H (2018) Biosynthesis of chitosan-Oligosaccharides (COS) by non-aflatoxigenic Aspergillus sp. strain EGY1 DSM 101520: a robust biotechnological approach. Process Biochem. 64: 16–30. https://doi.org/10.1016/j.procbio.2017.09.030

  38. Embaby AM, Melika RR, Hussein A, El-Kamel AH, Marey HS (2016) A novel non-cumbersome approach towards biosynthesis of pectic-oligosaccharides by non-aflatoxigenic Aspergillus sp. section flavi strain EGY1 DSM 101520 through citrus pectin fermentation. PLoS One 11:e0167981. https://doi.org/10.1371/journal.pone.0167981

    Article  Google Scholar 

  39. Kim H, Im Y, Choi J, Han SJ (2016) Optimization of physical factor for amylase production by Arthrobacter sp. by response surface methodology. Korean J Chem Eng 54:140–144

    Article  Google Scholar 

  40. Smith MR, Zahnley JC (2005) Production of amylase by Arthrobacter psychrolactophilus. J Ind Microbiol Biotechnol 32:277–283. https://doi.org/10.1007/s10295-005-0240-3

    Article  Google Scholar 

  41. Fan H, Liu Y, Liu Z (2009) [Optimization of fermentation conditions for cold-adapted amylase production by Micrococcus antarcticus and its enzymatic properties]. Huan jing ke xue= Huanjing kexue 30: 2473–2478.

  42. Cotârlet M, Negoiţă TG, Bahrim GE, Stougaard P (2011) Partial characterization of cold active amylases and proteases of Streptomyces sp. from Antarctica. Braz J Microbiol 42:868–877. https://doi.org/10.1590/S1517-83822011000300005

    Article  Google Scholar 

  43. Ji-lian WANG, Yu REN, Ming-yuan LI (2020) Medium optimization of cold-active amylase by Pseudomonas sp. using response surface methodology [J]. J Biol 37(4):101

    Google Scholar 

  44. Krishna C (2005) Solid-state fermentation systems-an overview. Crit Rev Biotechnol 25:1–30. https://doi.org/10.1080/07388550590925383

    Article  Google Scholar 

  45. Ottoni JR, Rodrigues e Silva T, Maia de Oliveira V, Zambrano Passarini MR

  46. Characterization of amylase produced by cold-adapted bacteria from Antarctic samples. Biocatal Agric Biotechnol 23: 101452

  47. Zhang J, Zeng R (2007) Psychrotrophic amylolytic bacteria from deep sea sediment of Prydz Bay, Antarctic: diversity and characterization of amylases. World J Microbiol Biotechnol 23:1551–1557. https://doi.org/10.1007/s11274-007-9400-0

    Article  Google Scholar 

  48. Arabacı N, Arıkan B (2018) Isolation and characterization of a cold-active, alkaline, detergent stable α-amylase from a novel bacterium Bacillus subtilis N8. Prep. Biochem Biotechnol 48:419–426. https://doi.org/10.1080/10826068.2018.1452256

    Article  Google Scholar 

  49. Liu J, Zhang Z, Liu Z, Zhu H, Dang H, Lu J, Cui Z (2011) Production of cold-adapted amylase by marine bacterium Wangia sp. C52: optimization, modeling, and partial characterization. Mar Biotechnol (NY) 13:837–844. https://doi.org/10.1007/s10126-010-9360-5

    Article  Google Scholar 

  50. Pandey A, Nigam P, Soccol CR, Soccol VT, Singh D, Mohan R (2000) Advances in microbial amylases. Biotechnol Appl Biochem 31:135–152. https://doi.org/10.1042/ba19990073

    Article  Google Scholar 

  51. Gupta R, Gigras P, Mohapatra H, Goswami VK, Chauhan B (2003) Microbial α-amylases: a biotechnological perspective. Process Biochem 38:1599–1616. https://doi.org/10.1016/S0032-9592(03)00053-0

    Article  Google Scholar 

  52. Qin Y, Huang Z, Liu Z (2014) A novel cold-active and salt-tolerant α-amylase from marine bacterium Zunongwangia profunda: molecular cloning, heterologous expression and biochemical characterization. Extremophiles 18:271–281. https://doi.org/10.1007/s00792-013-0614-9

    Article  Google Scholar 

  53. Rathour R, Gupta J, Tyagi B, Thakur I (2020) Production and characterization of psychrophilic α-amylase from a psychrophilic bacterium, Shewanella sp. ISTPL2. Amylase. https://doi.org/10.1515/amylase-2020-0001

  54. William Konigsberg, Reduction of disulfide bonds in proteins with dithiothreitol (1972) Methods in enzymology 25: 185-188

  55. Siddiqui KS, Poljak A, Guilhaus M, Feller G, D'Amico S, Gerday C, Cavicchioli R (2005) Role of disulfide bridges in the activity and stability of a cold-active α–amylase. J Bacteriol 187:6206–6212

    Article  Google Scholar 

  56. Jensen MT, Gottschalk TE, Svensson B (2003) Differences in conformational stability of barley alpha-amylase isozymes 1 and 2. Role of charged groups and isozyme 2 specific salt-bridges. J Cereal Sci 38:289–300

    Article  Google Scholar 

  57. Pons J, Planas A, Querol E (1995) Contribution of a disulfide bridge to the stability of 1,3-1,4-beta-D-glucan 4-glucanohydrolase from Bacillus licheniformis. Protein Eng 8:939–945. https://doi.org/10.1093/protein/8.9.939

    Article  Google Scholar 

  58. Otzen D (2011) Protein–surfactant interactions: a tale of many states. Biochim Biophys Acta - Proteins Proteomics 1814:562–591. https://doi.org/10.1016/j.bbapap.2011.03.003

    Article  Google Scholar 

  59. Wang X, Kan G, Shi C, Xie Q, Ju Y, Wang R, Qiao Y, Ren X (2019) Purification and characterization of a novel wild-type α-amylase from Antarctic sea ice bacterium Pseudoalteromonas sp. M175. Protein Expression and Purification 164:105444

    Article  Google Scholar 

  60. Xie F, Quan S, Liu D, Ma H, Li F, Zhou F, Chen G (2014) Purification and characterization of a novel α-amylase from a newly isolated Bacillus methylotrophicus strain P11-2. Process Biochem 49: 47–53. https://doi.org/10.1016/j.procbio.2013.09.025

  61. Fincan SA, Enez B, Özdemir S, Bekler FM (2014) Purification and characterization of thermostable α-amylase from thermophilic Anoxybacillus flavithermus. Carbohydrate polymers 102:144–150

    Article  Google Scholar 

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  1. Institute of Graduate Studies and Research, Environmental Studies Department, Alexandria University, Alexandria, Egypt

    Amira A. Matrawy & Ahmed I. Khalil

  2. Institute of Graduate Studies and Research, Biotechnology Department, Alexandria University, 163 Horreya Avenue, Chatby, Alexandria, 21526, Egypt

    Amira M. Embaby

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  1. Amira A. Matrawy
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  2. Ahmed I. Khalil
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AK: supervision, conceptualization, and revising of the manuscript. AE: supervision, methodology, conceptualization, data curation, and writing — reviewing and editing the manuscript. AM: visualization, investigation, validation, data curation, and writing — original draft preparation.

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Correspondence to Amira M. Embaby.

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Fig. S1

Eadie-Hofstee plot of purified CA-AM21 using soluble starch as a substrate. The plot was drawn by Hyper32 software (PNG 37 kb)

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Matrawy, A.A., Khalil, A.I. & Embaby, A.M. Bioconversion of bread waste by marine psychrotolerant Glutamicibacter soli strain AM6 to a value-added product: cold-adapted, salt-tolerant, and detergent-stable α-amylase (CA-AM21). Biomass Conv. Bioref. 13, 12125–12142 (2023). https://doi.org/10.1007/s13399-022-02325-3

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