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. 2023 Apr 3;12(7):1499.
doi: 10.3390/foods12071499.

Production of a Series of Long-Chain Isomaltooligosaccharides from Maltose by Bacillus subtilis AP-1 and Associated Prebiotic Properties

Suratsawadee Tiangpook  1 , Sreyneang Nhim  1 , Pattaneeya Prangthip  2 , Patthra Pason  1   3 , Chakrit Tachaapaikoon  1   3 , Khanok Ratanakhanokchai  1   3 , Rattiya Waeonukul  1   3
Affiliations

Production of a Series of Long-Chain Isomaltooligosaccharides from Maltose by Bacillus subtilis AP-1 and Associated Prebiotic Properties

Suratsawadee Tiangpook et al. Foods. .

Abstract

Bacillus subtilis strain AP-1, which produces α-glucosidase with transglucosidase activity, was used to produce a series of long-chain isomaltooligosaccharides (IMOs) with degree of polymerization (DP) ranging from 2 to 14 by direct fermentation of maltose. A total IMOs yield of 36.33 g/L without contabacillusmination from glucose and maltose was achieved at 36 h of cultivation using 50 g/L of maltose, with a yield of 72.7%. IMOs were purified by size exclusion chromatography with a Superdex 30 Increase column. The molecular mass and DP of IMOs were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). Subsequently, linkages in produced oligosaccharides were verified by enzymatic hydrolysis with α-amylase and oligo-α-1,6-glucosidase. These IMOs showed prebiotic properties, namely tolerance to acidic conditions and digestive enzymes of the gastrointestinal tract, stimulation of probiotic bacteria growth to produce short-chain fatty acids and no stimulating effect on pathogenic bacteria growth. Moreover, these IMOs were not toxic to mammalian cells at up to 5 mg/mL, indicating their biocompatibility. Therefore, this research demonstrated a simple and economical method for producing IMOs with DP2-14 without additional operations; moreover, the excellent prebiotic properties of the IMOs offer great prospects for their application in functional foods.

Keywords: Bacillus subtilis; isomaltooligosaccharide; prebiotic; transglucosidase; α-glucosidase.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
TLC patterns of culture supernatant of B. subtilis strain AP-1 grown in MS medium containing 5% (w/v) maltose. Lane S1: glucose (G), isomaltose (IMO2) and isomaltotriose (IMO3) standard; lane S2: maltose (M2) standard.
Figure 2
Figure 2
MALDI-TOF-MS spectra of oligosaccharides obtained from the culture supernatant of B. subtilis strain AP-1 at 36 h.
Figure 3
Figure 3
(A) TLC patterns of elution profile of IMOs obtained from the culture supernatant of B. subtilis strain AP-1 at 36 h using a Superdex 30 Increase column with a flow rate of 0.1 mL/min. (B,C) TLC patterns of elution profiles of samples F1 and F2, respectively, using a Superdex 30 Increase column with a flow rate of 0.025 mL/min. Lane Std: glucose (G), isomaltose (IMO2) and isomaltotriose (IMO3) standard; lane M: IMO mixture from culture supernatant at 36 h of B. subtilis strain AP-1; lane F1: sample F1; lane F2: sample F2.
Figure 4
Figure 4
MALDI-TOF-MS spectra of purified IMOs produced from B. subtilis strain AP-1 with degree of polymerization (DP) as follows: (A) DP2, (B) DP3, (C) DP4, (D) DP5, (E) DP6, (F) DP7, (G) DP8, (H) DP9 and (I) DP10.
Figure 5
Figure 5
TLC of enzymatic hydrolysis of maltohexaose (M6) (A) and each IMO with degree of polymerization (DP) 2–10 produced from B. subtilis strain AP-1 (B) by the action of α-amylase (AM) and oligo-α-1,6-glucosidase (α1,6). Glucose (G), maltooligosaccharides (M2–M6), isomaltose (IMO2) and isomaltotriose (IMO3) were used as standards (shown on the left-hand side of each image).
Figure 6
Figure 6
Effect of oligosaccharides on the cell viability of mammalian cells. Data are the means of three independent experiments and error bars represent the standard deviation. IMOs, isomaltooligosaccharides; FOSs, fructooligosaccharides.
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