Mass Spectrometry-Based Metabolomics of Agave Sap (Agave salmiana) after Its Inoculation with Microorganisms Isolated from Agave Sap Concentrate Selected to Enhance Anticancer Activity

Luis Figueroa,Janet Gutiérrez-Uribe,Liliana Santos-Zea,Adelfo Escalante

doi:10.3390/su9112095

Luis Figueroa, Janet Gutiérrez-Uribe + Show 2 more

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https://doi.org/10.3390/su9112095

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Abstract

Saponins have been correlated with the reduction of cancer cell growth and the apoptotic effect of agave sap concentrate. Empirical observations of this artisanal Mexican food have shown that fermentation occurs after agave sap is concentrated, but little is known about the microorganisms that survive after cooking, or their effects on saponins and other metabolites. The aim of this study was to evaluate the changes in metabolites found in agave (A. salmiana) sap after its fermentation with microorganisms isolated from agave sap concentrate, and demonstrate its potential use to enhance anticancer activity. Microorganisms were isolated by dilution plating and identified by 16S rRNA analysis. Isolates were used to ferment agave sap, and their corresponding butanolic extracts were compared with those that enhanced the cytotoxic activity on colon (Caco-2) and liver (Hep-G2) cancer cells. Metabolite changes were investigated by mass spectrometry-based metabolomics. Among 69 isolated microorganisms, the actinomycetes Arthrobacter globiformis and Gordonia sp. were used to analyze the metabolites, along with bioactivity changes. From the 939 ions that were mainly responsible for variation among fermented samples at 48 h, 96 h, and 192 h, four were correlated to anticancer activity. It was shown that magueyoside B, a kammogenin glycoside, was found at higher intensities in the samples fermented with Gordonia sp. that reduced Hep-G2 viability better than controls. These findings showed that microorganisms from agave sap concentrate change agave sap metabolites such as saponins. Butanolic extracts obtained after agave sap fermentation with Arthrobacter globiformis or Gordonia sp. increased the cancer cell growth inhibitory effect on colon or liver cancer cells, respectively.

Highlights

Agave sap concentrate is a food produced by thermal treatment of the fresh sap (“aguamiel”)
Microorganisms Isolation, Screening, and Identification Agave sap microorganisms were mainly grown in R2A and NZ amine A media with 38 and 29 isolatesA, graevspe escatpivmeilcyr.oNorogagnrioswmtshwweraesmdaeitnelcytegdroiwnnthine RM2RASamndeNdiZumam. iOnenlAy monedeiiasowliatthe3w8 aans dde2t9ected in YMiYsMoalnaadtensPd, DrPeADspAmecmteidvedeialiya, .,wNwhohiicgchrhowwwatahss wiinnaddsiicdcaaetttievivceteethdthaitanttthhteheepMrpeRrsSeensmecneedcoeifuoymef.ayOsetnsalsiyntsotnihnee ticshooenlaccteoennwtrcaaetsnedtiresateltiecmtieisdtelidinm. ited
Microorganisms that survive the thermal treatment of agave sap have the potential to modify agave sap metabolites such as saponins

Summary

Introduction

Agave sap concentrate is a food produced by thermal treatment of the fresh sap (“aguamiel”). Agave sap is composed of water, sugars (glucose, fructose, and sucrose), proteins, gums, and mineral salts [1] It is obtained from the mature plant, which is between eight and 10 years old. Saponins have been isolated mainly from the leaves of the plant [4], but recently they were reported in agave sap concentrate [5,6], and fresh agave sap [7]. These molecules were identified as glycosides of kammogenin, manogenin, gentrogenin, and hecogenin, and are the main source of variability among agave sap concentrates [6]. A kammogenin glycoside identified as magueyoside B has been correlated with reduction in viability, as well as the induction of apoptosis in colon cancer cell lines [5,6]

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