Mushroom bioreactor-biomass as bioactive protein source: Synergy of mushroom rural and urban cultivation

doi:10.21203/rs.3.rs-3332222/v1

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Mushroom bioreactor-biomass as bioactive protein source: Synergy of mushroom rural and urban cultivation

https://doi.org/10.21203/rs.3.rs-3332222/v1

This work is licensed under a CC BY 4.0 License

Journal Publication

published 12 Feb, 2024

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Mushroom rural cultivation consumes averagely 6-month period, while urban cultivation takes only 10 days or less. In this study, mushroom biomass was grinded and converted into a flour to produce mushroom-chicken patties using Lingzhi and Enoki. The inclusion of Enoki in chicken patties (10%, 20% and 30%) indicates higher consumer acceptance significantly (p > 0.05) compared to chicken patties with Lingzhi (10% and 20%). This analysis validated the concept of mushroom biomass as source of bioactive protein. On the other hand, 3kg dried mushroom-bioreactor biomass was produced using a heterotrophic 1m² fabricated-bioreactor, which answers the minimum requirement for protein content for 1 human per year. Together, these explain the significance of mushroom biomass in food security as a protein source and the synergy of mushroom rural-urban cultivation.

Mushroom biomass

Bioreactor

Protein

Urban cultivation

Landless food

The global wheat flour’s price soars as a result of Russian’s invasion of Ukraine, one of the major producers of wheat and heatwave in India, second global wheat producer, causing the crops’ withering (Vethasalam, 2022). With the current concerning issues, wheat flour’s shortage and occurrence of land’s heatwaves, researchers are looking into solutions to generate food using landless environment with lesser time and cost consumption. The production of food involves growing, cooking, distributing, transporting, disposing, and consuming. For decades to come, there will likely be an increase in the world's food demand, which could lead to both agricultural reinforcement and growth (Olén et al., 2021). The global food system is responsible for around one-fourth of all greenhouse gas (GHG) emissions. More than 50% of the methane (CH₄) and 75% of the nitrous oxide (N₂O) emissions from human activities globally generated from crop and livestock production within the farm gate (FAO, 2020). The severity of these significant consequences is increased by emissions from pre- and post-production activities such as the production of fertiliser, food transport, processing, retail, and waste disposal (Tubiello et al., 2021).

Besides that, consumers concern about health and sustainability are driving changes from land use to ingredient supply chain. In addition, technology advancements are reshaping globally, and mushroom-bioreactor technology is designed to help to achieve higher yields and productivity without compromising mushroom’s nutrients properties. Mushroom proteins have recently attracted the interest of the scientific community and participants in the food industry due to their excellent nutritional value and high concentration of key amino acids when compared to vegetables (González et al., 2021). Besides that, mushrooms also exhibit a great level of peptides and proteins with natural lectins, fungal immunomodulatory proteins, ribosome inactivating proteins, antimicrobial proteins, ribonucleases, and laccases (Valverde et al., 2015).

In recent studies, mushroom biomass has been used successfully as functional enhancers in various food products (Wan-Mohtar et al., 2018; Wan-Mohtar et al., 2020). In order to achieve fast cultivation time, the development of controlled cultivation has proven to produce mushroom biomass in 10 days or less (Supramani et al., 2019). Bioreactor-grown mushroom biomass, particularly Lingzhi, has been accepted as an alternative animal feed due to its great nutrients value such as protein (32.23%), dietary fibre (13.8%), carbohydrate (48.38%), ash (1.14%) and lipids (4.45%) contents (Wan-Mohtar et al., 2021). The high protein content warranted of using this biomass as a ‘’Novel Food’’ development for human consumption.

Besides these great nutrients, mushroom has also been proven as functional food for their nutraceuticals properties including anti-inflammatory, anticarcinogenic, antitumor, antimutagenic, antidiabetic, antibacterial, antiviral, anti-obesity, and anti-hypercholesterolemic activities (Das et al., 2021).

The radar chart showed in Fig. 1 summarizes the panellists’ scores for each attribute. The commercial chicken patties collectively obtained the highest scores in all sensory attributes. Regarding the chicken patties formulated with different concentration of mushroom flour, the 10% Enoki-chicken patty (6.73) showed a great acceptance by the panellists, and significant appearance (7.17), colour (7.20) and aroma (6.93) than the control chicken patty (6.73, 6.83, 6.60 respectively). Also, the addition of 10% Lingzhi in chicken patty scores (3.03) in overall acceptability, compared to chicken patties with Lingzhi (20%) obtained the least attributes and acceptability scores.

Preliminary study

Both fruiting bodies of Lingzhi and Enoki mushrooms were dried to prepare mushroom biomass. The biomass was grinded and turned into mushroom flour (MF) as shown in Fig. 1. Based on previous study by Wan-Mohtar et al. (2020), the formulated mushroom-chicken patty samples; 10%, 20% and 30% Enoki, 10% and 20% Lingzhi, control and commercial chicken patty were assessed for appearance, colour, aroma, texture, taste, aftertaste and overall acceptability using a 9-point hedonic scale (1 = extremely poor, 2 = very poor, 3 = poor, 4 = bad, 5 = average, 6 = fair, 7 = good, 8 = very good, 9 = excellent), by 30 acceptable panellists from Biotechnology Program, Universiti Malaya, Kuala Lumpur, Malaysia. The samples were distributed to each of the panellist, together with a cup of drinking water and a piece of unsalted cracker for the purpose of palate-cleansing during the evaluation.

Table 1 depicts the overall acceptability scores of utilisation of Lingzhi and Enoki mushroom in chicken patty and other addition of mushroom species in meat products. Chicken patty with Enoki mushroom shows higher acceptance compared to pork sausage with 1% winter mushroom (5.46). Besides that, both addition of 30% Agaricus and 2% straw mushrooms in sausages shows prominent acceptance scores of 9.50 and 9.00 respectively. However, 45% PSC mushroom-chicken sausage scores 5.20 of overall acceptability, and 10% FBB chicken patty exhibits overall acceptability of 6.0. Although both meat products presented significant scores, they produced lower scores compared to 10% Enoki mushroom -chicken patty by this study.

The incorporation of Enoki mushroom flour in the development of chicken patties showed a greater potency, whereas the chicken patties formulated with Lingzhi mushroom flour were less acceptable to meet the consumer’s palate due to its strong unique flavour and an undesirable appearance in comparison to commercial chicken patty.

Mushroom biomass can be produced by both rural and urban cultivation (Fig. 2). Rural production requires a big arable land in order to grow crops and accommodate a vast number of mushroom bags. Besides that, the production of rural mushroom, for example, Ganoderma lucidum requires a longer time (6 months) compared to urban cultivation (10 days or less) (Hanafiah et al., 2019). Due to extensive fermentation period, risk of contamination of solid state fermentation (SSF) of rural cultivation is higher compared to submerged liquid fermentation (SLF) due to quality and environment of culture (Hassan et al., 2019). During post-harvest, rural mushroom experiences extreme textural changes mainly, loss of firmness or softening due to membrane alterations and hyphae shrinking, polysaccharide and protein degradation, central vacuole disruption, and intercellular space expansion at the pileus surface (Wan-Mohtar et al., 2019). Meanwhile, an effective production of mycelial pellets or mushroom-bioreactor biomass by heterotrophic 1m² bioreactor has been proven by Usuldin et al. (2021). In addition, low contamination risks from biological and chemical factors are also an added element of bioreactor systems to a successful biomass cultivation (Bakratsas et al., 2021). Due to the stability of growth parameters, efficient use of nutrients (which can be supplied precisely in accordance with demand), low water and land footprint, and lack of pesticides or antibiotics, producing biomass in a bioreactor is significantly more efficient than growing plants or raising animals in an open field (Ciani et al., 2021). SLF of urban cultivation has become an innovative and significant method for obtaining the bioactive ingredients of Ganoderma lucidum given its rapid production cycle, controlled procedure, high output, consistent product quality, and cost-effectiveness (Klaus & Wan-Mohtar, 2022).

The strategy, which in accordance with landless food concept by Rahmann et al. (2020), is to apply mushroom urban production to produce mushroom-bioreactor biomass for human consumption. Based on Fig. 3, 3kg of biomass can be produced in 1 run resulting 0.966kg of protein. The protein value of biomass is calculated using ratio of the protein value (g/100g) by Wan-Mohtar et al. (2021). Furthermore, this mushroom-bioreactor biomass not only contain high protein value, but also consists of other vital nutrients for balanced diet such as carbohydrate, fibre and lipid (Wan-Mohtar et al., 2021). In addition, the primary nutrients from animal manure are nitrogen, phosphorus and potassium. Despite the importance of organic fertilisers in agricultural output, animal manure is a valuable source of soil fertiliser, and contribute to conversion to food and energy (Chew et al., 2019). Besides manure, mushroom also contributes in enhancing crops production (Ferraro et al., 2022).

Currently, mushrooms are actively utilized in the industries of fermentation, food, pharmacy, and medicine. Applying plant-based resources such as mushroom is more fundamentally ethical and respects the welfare of animals (Nasrabadi et al., 2021). Therefore, consumers and environmental concerns are also driving the transition away from animal-based resources and towards plant-based ones. However, there are numerous key obstacles to mushroom-bioreactor biomass food need to be given consideration:

Consumer-based

Firstly, human diet has included meat from the early days, and it has vital cultural and culinary significance. Unsurprisingly, a lot of individuals believe that meat is an essential component of the meal from a cultural and nutritional standpoint. Meat has also been linked to more formal meals like restaurant dinners or corporate meals, where it is thought to be more acceptable than meat alternatives (Michel et al., 2021). When consumers first experiment with plant-based substitutes, they frequently experience food neophobia, which is defined by a fear of eating novel or unfamiliar foods (Tuorila & Hartmann, 2020).

Sensory and technology-based

There are nonetheless numerous issues that need to be resolved despite the adoption of several techniques that change the structure of plant proteins or raise their capacity to hold water. A distinctive juiciness quality of meat brought on by water absorption and the way that water binds to proteins and their fibres (Kurek et al., 2022). Moreover, meat products are characterised by a red, reddish, or pink colour that is evidently challenging to achieve without the application of colourants, in addition to texture and palatability issues (McClements & Grossmann, 2021).

Commercial-based

Since they feature numerous claims that tend to highlight the qualities of organic, vegan, and non-GMO products that give rise to the image of ‘healthy food’, the products had a surprising amount of additives (D’Alessandro et al., 2022). Preservatives, stabilisers, colourants, and thickeners are common additives in vegetarian goods that serve as meat protein replacements (Sha & Xiong, 2020). Besides, the price of plant proteins (in comparison to those from animal sources) also has come under intense debate as a result of the rise in popularity of plant-based foods (Fehér et al., 2020).

Current prospects and future work

The pressing crises of the current days and future, such as competition for natural resources, population expansion, food losses and waste, climate change cause a danger to the current agriculture and food systems. Due to these factors, a change and exposure to more sustainable food production methods and products is necessary for our agricultural landscape. Furthermore, greater emphasis is needed for the development of various functional food and other potential products using mushroom-bioreactor biomass from urban cultivation for future food and long-term prospects. Increasing mushroom production appears to be a feasible and alluring solution for more sustainable farming on a worldwide scale. Also, enhancing the economic worth of mushroom, whether they are in their raw or processed state, may concentrate demand and promote market orientation.

In this study, synergism of mushroom rural to urban cultivation has been achieved. Lingzhi and Enoki mushroom biomass significantly affected the texture, colour, taste, aftertaste, and overall acceptability of the chicken patties. Notably, 10% Enoki-chicken patty indicates moderately higher consumer acceptability score (6.73) compared to chicken patties with 10% and 20% Lingzhi mushroom (3.03 and 2.10 respectively). In addition, 3kg mushroom-bioreactor biomass with 32.2% protein has been obtained using heterotrophic 1m² bioreactor. By using technology to lower cultivation startup costs and to enhance the quality and productivity of each mushroom growth cycle, mushroom cultivating can change the economic, social, and environmental norm in low- and middle-income countries. Therefore, with the present worldwide concerns of flour shortages and land heatwaves, this rapid and sustainable mushroom production will potentially promote the utilisation of mushroom-bioreactor biomass as future bioactive protein source.

No conflict interest.

Funding Declaration

The research is sponsored by Ministry of Higher Education (MOHE), Malaysia

Author Contribution

Visualization, N.A.Z.A.; Supervision, W.A.A.Q.I.W.-M. AND A.A.J.; Project Administration, W.A.A.Q.I.W.-M.; Funding Acquisition, W.A.A.Q.I.W.-M,; Resources, W.A.A.Q.I.W.-M.; Data Curation, N.A.Z.A. and W.A.A.Q.I.W.-M.Conceptualization, W.A.A.Q.I.W.-M., Z.I., N.A.Z.A, T.R.Y. and A.A.J.; Methodology, N.A.Z.A. ; Formal Analysis, T.R.Y.; Investigation, W.A.A.Q.I.W.-M.;; Writing—Original Draft Preparation, N.A.Z.A. and W.A.A.Q.I.W.-M.; Writing—Review and Editing, N.A.Z.A.;. All authors have read and agreed to the published version of the manuscript.

Acknowledgements

I would like to thank the Ministry of Higher Education (MOHE) under the Fundamental Research Grant Scheme (FRGS:FP062-2022) (FRGS/1/2022/STG01/UM/02/2) for this project.

Data Availability

The raw data will be provide upon request

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Table 1

Overall acceptability of meat products utilising different mushroom species
Mushroom species	Meat product	Concentration	Function	Overall acceptability	Reference
Lingzhi and Enoki mushrooms	Chicken patty	10%, 20%, 30%	Functional flour	3.03 (10% Lingzhi) and 6.73 (10% Enoki)	This study
Winter mushroom (Flammulina velutipes)	Emulsion type pork sausage	0, 0.5%, 1.0%, 1.5%, and 2.0%	Natural alternative to phosphates.	5.46 (1.0%)	(Choe et al., 2018)
Agaricus spp.	Beef sausage	0, 15%, 30% and 45%	Meat substitution	9.50 (30%)	(Al-Dalain, 2018)
Straw mushrooms (Volvariella volvacea)	Cantonese sausage	0%, 1%, 2%, 3%, and 4%	Amino acids and volatile compounds enhancer, and peroxide reducer	9.00 (2%)	(Wang et al., 2018)
Pleurotus sajur-caju (PSC) mushroom	Chicken sausage	0, 15%, 30%, 45% and 60%	Fat-lowering and fibre-improving agents	5.20 (45%)	(Yahya & Ting, 2020)
Oyster mushroom fruiting-body-base (FBB)	Chicken patty	10%, 20%, 30%	Antioxidative flour	6.00 (10%)	(Wan-Mohtar et al., 2020)

No competing interests reported.

Download PDF

Journal Publication

published 12 Feb, 2024

Read the published version in Organic Agriculture →

Editorial decision: Accepted
26 Jan, 2024
Reviews received at journal
27 Oct, 2023
Reviewers agreed at journal
18 Oct, 2023
Reviewers invited by journal
18 Oct, 2023
Editor assigned by journal
21 Sep, 2023
Submission checks completed at journal
13 Sep, 2023
First submitted to journal
06 Sep, 2023

You are reading this latest preprint version

Mushroom bioreactor-biomass as bioactive protein source: Synergy of mushroom rural and urban cultivation

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Abstract

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Discussion

Challenges, current prospects and future work

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